Isolated and fixed micro and nano structures and methods thereof

ABSTRACT

Discrete micro and nanoscale particles are formed in predetermined shapes and sizes and predetermined size dispersions. The particles can also be attached to a film to form arrays of particles on a film. The particles are formed from molding techniques that can include high throughput and continuous particle molding.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is based on and claims priority of U.S. ProvisionalPatent Application Ser. No. 60/734,228, filed Nov. 7, 2005; U.S.Provisional Patent Application Ser. No. 60/762,802, filed Jan. 27, 2006;and U.S. Provisional Patent Application No. 60/799,876, filed May 12,2006; the disclosure of each of which is incorporated herein byreference in its entirety.

This application is also a continuation-in-part of PCT InternationalApplication Serial No. PCT/US06/23722, filed Jun. 19, 2006, which isbased on and claims priority to U.S. Provisional Patent Application Ser.No. 60/691,607, filed Jun. 17, 2005; United States Provisional PatentApplication Ser. No. 60/714,961, filed Sep. 7, 2005; U.S. ProvisionalPatent Application Ser. No. 60/762,802, filed Jan. 27, 2006; and U.S.Provisional Patent Application No. 60/799,876, filed May 12, 2006; eachof which is incorporated herein by reference in its entirety.

This application is also a continuation-in-part of PCT InternationalApplication Serial No. PCT/US06/34997, filed Sep. 7, 2006, which isbased on and claims priority to U.S. Provisional Patent Application Ser.No. 60/714,961, filed Sep. 7, 2005; U.S. Provisional Patent ApplicationSer. No. 60/734,228, filed Nov. 7, 2005; U.S. Provisional PatentApplication Ser. No. 60/762,802, filed Jan. 27, 2006; and U.S.Provisional Patent Application No. 60/799,876, filed May 12, 2006; eachof which is incorporated herein by reference in its entirety.

This application is also a continuation-in-part of U.S. patentapplication Ser. No. 10/583,570, filed Jun. 19, 2006, which is based onand claims priority to PCT International Patent Application Serial No.PCT/US04/42706, filed Dec. 20, 2004, which is based on and claimspriority to U.S. Provisional Patent Application Ser. No. 60/531,531,filed Dec. 19, 2003; U.S. Provisional Patent Application Ser. No.60/583,170, filed Jun. 25, 2004; and U.S. Provisional Patent ApplicationSer. No. 60/604,970, filed Aug. 27, 2004, each of which is incorporatedherein by reference in its entirety.

GOVERNMENT INTEREST

A portion of the disclosure contained herein was made with U.S.Government support from the Office of Naval Research Grant No.N00014210185 and the Science and Technology Center program of theNational Science Foundation under Agreement No. CHE-9876674. The U.S.Government has certain rights to that portion of the disclosure.

INCORPORATION BY REFERENCE

All documents referenced herein are hereby incorporated by reference asif set forth in their entirety herein.

TECHNICAL FIELD

Generally, the present disclosure relates to micro and nanoscaleparticles and methods for forming the particles.

ABBREVIATIONS

° C.=degrees Celsius

cm=centimeter

DBTDA=dibutyltin diacetate

DMA=dimethylacrylate

DMPA=2,2-dimethoxy-2-phenylacetophenone

EIM=2-isocyanatoethyl methacrylate

FEP=fluorinated ethylene propylene

Freon 113=1,1,2-trichlorotrifluoroethane

g=grams

h=hours

Hz=hertz

IL=imprint lithography

kg=kilograms

kHz=kilohertz

kPa=kilopascal

MCP=microcontact printing

MEMS=micro-electro-mechanical system

MHz=megahertz

MIMIC=micro-molding in capillaries

mL=milliliters

mm=millimeters

mmol=millimoles

mN=milli-Newton

m.p.=melting point

mW=milliwatts

NCM=nano-contact molding

NIL=nanoimprint lithography

nm=nanometers

PDMS=polydimethylsiloxane

PEG poly(ethylene glycol)

PFPE=perfluoropolyether

PLA poly(lactic acid)

PP=polypropylene

Ppy=poly(pyrrole)

psi=pounds per square inch

PVDF=poly(vinylidene fluoride)

PTFE=polytetrafluoroethylene

SAMIM=solvent-assisted micro-molding

SEM=scanning electron microscopy

S-FIL=“step and flash” imprint lithography

Si=silicon

Tg=glass transition temperature

Tm=crystalline melting temperature

TMPTA=trimethylolpropane triacrylate

μM=micrometers

UV=ultraviolet

w=watts

ZDOL=poly(tetrafluoroethylene oxide-co-difluoromethylene oxide)α,ω diol

BACKGROUND

The availability of viable nanofabrication processes is a key factor torealizing the potential of nanotechnologies. In particular, theavailability of viable nanofabrication processes is important to thefields of photonics, electronics, and proteomics. Traditional imprintlithographic (IL) techniques are an alternative to photolithography formanufacturing integrated circuits, micro- and nano-fluidic devices, andother devices with micrometer and/or nanometer sized features. There isa need in the art, however, for new materials to advance IL techniques.See Xia, Y., et al., Angew. Chem. Int. Ed., 1998, 37, 550-575; Xia, Y.,et al., Chem. Rev., 1999, 99, 1823-1848; Resnick, D. J., et al.,Semiconductor International, June, 2002, 71-78; Choi, K. M., et al., J.Am. Chem. Soc., 2003, 125, 40604061; McClelland, G. M., et al., Appl.Phys. Lett., 2002, 81, 1483; Chou, S. Y., et al., J. Vac. Sci. Technol.B, 1996, 14, 4129; Otto, M., et al., Microelectron. Eng., 2001, 57, 361;and Bailey, T., et al., J. Vac. Sci. Technol., B, 2000, 18, 3571.

Imprint lithography includes at least two areas: (1) soft lithographictechniques, see Xia, Y., et al., Angew. Chem. Int. Ed., 1998, 37,550-575, such as solvent-assisted micro-molding (SAMIM); micro-moldingin capillaries (MIMIC); and microcontact printing (MCP); and (2) rigidimprint lithographic techniques, such as nano-contact molding (NCM), seeMcClelland, G. M., et al., Appl. Phys. Lett., 2002, 81, 1483; Otto, M.,et al., Microelectron. Eng., 2001, 57, 361; “step and flash” imprintlithographic (S-FIL), see Bailey, T., et al., J. Vac. Sci. Technol., B,2000, 18, 3571; and nanoimprint lithography (NIL), see Chou, S. Y., etal., J. Vac. Sci. Technol. B, 1996, 14, 4129.

Polydimethylsiloxane (PDMS) based networks have been the material ofchoice for much of the work in soft lithography. See Quake, S. R., etal., Science, 2000, 290, 1536; Y. N. Xia and G. M. Whitesides, Angew.Chem. Int Ed. Engl. 1998, 37, 551; and Y. N. Xia, et al., Chem. Rev.1999, 99, 1823.

The use of soft, elastomeric materials, such as PDMS, offers severaladvantages for lithographic techniques. For example, PDMS is highlytransparent to ultraviolet (UV) radiation and has a very low Young'smodulus (approximately 750 kPa), which gives it the flexibility requiredfor conformal contact, even over surface irregularities, without thepotential for cracking. In contrast, cracking can occur with molds madefrom brittle, high-modulus materials, such as etched silicon and glass.See Bietsch, A., et al., J. Appl. Phys., 2000, 88, 43104318. Further,flexibility in a mold facilitates the easy release of the mold frommasters and replicates without cracking and allows the mold to enduremultiple imprinting steps without damaging fragile features.Additionally, many soft, elastomeric materials are gas permeable, aproperty that can be used to advantage in soft lithography applications.

Although PDMS offers some advantages in soft lithography applications,several properties inherent to PDMS severely limit its capabilities insoft lithography. First, PDMS-based elastomers swell when exposed tomost organic soluble compounds. See Lee, J. N., et al., Anal. Chem.,2003, 75, 6544-6554. Although this property is beneficial inmicrocontact printing (MCP) applications because it allows the mold toadsorb organic inks, see Xia, Y., et al., Angew. Chem. Int. Ed., 1998,37, 550-575, swelling resistance is critically important in the majorityof other soft lithographic techniques, especially for SAMIM and MIMIC,and for IL techniques in which a mold is brought into contact with asmall amount of curable organic monomer or resin. Otherwise, thefidelity of the features on the mold is lost and an unsolvable adhesionproblem ensues due to infiltration of the curable liquid into the mold.Such problems commonly occur with PDMS-based molds because most organicliquids swell PDMS. Organic materials, however, are the materials mostdesirable to mold. Additionally, acidic or basic aqueous solutions reactwith PDMS, causing breakage of the polymer chain.

Secondly, the surface energy of PDMS (approximately 25 mN/m) is not lowenough for soft lithography procedures that require high fidelity. Forthis reason, the patterned surface of PDMS-based molds is oftenfluorinated using a plasma treatment followed by vapor deposition of afluoroalkyl trichlorosilane. See Xia, Y., et al., Angew. Chem. Int. Ed.,1998, 37, 550-575. These fluorine-treated silicones swell, however, whenexposed to organic solvents.

Third, the most commonly-used commercially available form of thematerial used in PDMS molds, e.g., Sylgard 184® (Dow CorningCorporation, Midland, Mich., United States of America) has a modulusthat is too low (approximately 1.5 MPa) for many applications. The lowmodulus of these commonly used PDMS materials results in sagging andbending of features and, as such, is not well suited for processes thatrequire precise pattern placement and alignment. Although researchershave attempted to address this last problem, see Odom, T. W., et al., J.Am. Chem. Soc., 2002, 124, 12112-12113; Odom, T. W. et al., Langmuir,2002, 18, 5314-5320; Schmid, H., et al., Macromolecules, 2000, 33,3042-3049; Csucs, G., et al., Langmuir, 2003, 19, 6104-6109; Trimbach,D., et al., Langmuir, 2003, 19, 10957-10961, the materials chosen stillexhibit poor solvent resistance and require fluorination steps to allowfor the release of the mold.

Rigid materials, such as quartz glass and silicon, also have been usedin imprint lithography. See Xia, Y., et al., Angew. Chem. Int Ed., 1998,37, 550-575; Resnick, D. J., et al., Semiconductor International, June,2002, 71-78; McClelland, G. M., et al., Appl. Phys. Left., 2002, 81,1483; Chou, S. Y., et al., J. Vac. Sci. Technol. B, 1996, 14, 4129;Otto, M., et al., Microelectron. Eng., 2001, 57, 361; and Bailey, T., etal., J. Vac. Sci. Technol., B, 2000, 18, 3571; Chou, S. Y., et al.,Science, 1996, 272, 85-87; Von Werne, T. A., et al., J. Am. Chem. Soc.,2003, 125, 3831-3838; Resnick, D. J., et al., J. Vac. Sci. Technol. B,2003, 21, 2624-2631. These materials are superior to PDMS in modulus andswelling resistance, but lack flexibility. Such lack of flexibilityinhibits conformal contact with the substrate and causes defects in themask and/or replicate during separation.

Another drawback of rigid materials is the necessity to use a costly anddifficult to fabricate hard mold, which is typically made by usingconventional photolithography or electron beam (e-beam) lithography. SeeChou, S. Y., et al., J. Vac. Sci. Technol. B, 1996, 14, 4129. Morerecently, the need to repeatedly use expensive quartz glass or siliconmolds in NCM processes has been eliminated by using an acrylate-basedmold generated from casting a photopolymerizable monomer mixture againsta silicon master. See McClelland, G. M., et al., Appl. Phys. Lett.,2002, 81, 1483, and Jung, G. Y., et al., Nanoletters, 2004, ASAP. Thisapproach also can be limited by swelling of the mold in organicsolvents.

Despite such advances, other disadvantages of fabricating molds fromrigid materials include the necessity to use fluorination steps to lowerthe surface energy of the mold, see Resnick, D. J., et al.,Semiconductor International, June, 2002, 71-78, and the inherent problemof releasing a rigid mold from a rigid substrate without breaking ordamaging the mold or the substrate. See Resnick, D. J., et al.,Semiconductor International, June, 2002, 71-78; Bietsch, A, J. Appl.Phys., 2000, 88, 4310-4318. Khang, D. Y., et al., Langmuir, 2004, 20,2445-2448, have reported the use of rigid molds composed of thermoformedTeflon AF® (DuPont, Wilmington, Del., United States of America) toaddress the surface energy problem. Fabrication of these molds, however,requires high temperatures and pressures in a melt press, a process thatcould be damaging to the delicate features on a silicon wafer master.Additionally, these molds still exhibit the intrinsic drawbacks of otherrigid materials as outlined hereinabove.

Further, a clear and important limitation of fabricating structures onsemiconductor devices using molds or templates made from hard materialsis the usual formation of a residual or “scum” layer that forms when arigid template is brought into contact with a substrate. Even withelevated applied forces, it is very difficult to completely displaceliquids during this process due to the wetting behavior of the liquidbeing molded, which results in the formation of a scum layer. Thus,there is a need in the art for a method of fabricating a pattern or astructure on a substrate, such as a semiconductor device, which does notresult in the formation of a scum layer.

The fabrication of solvent resistant, microfluidic devices with featureson the order of hundreds of microns from photocurable perfluoropolyether(PFPE) has been reported. See Rolland, J. P., et al., J. Am. Chem. Soc.,2004, 126, 2322-2323. PFPE-based materials are liquids at roomtemperature and can be photochemically cross-linked to yield tough,durable elastomers. Further, PFPE-based materials are highly fluorinatedand resist swelling by organic solvents, such as methylene chloride,tetrahydrofuran, toluene, hexanes, and acetonitrile among others, whichare desirable for use in microchemistry platforms based on elastomericmicrofluidic devices. There is a need in the art, however, to applyPFPE-based materials to the fabrication of nanoscale devices for relatedreasons.

Further, there is a need in the art for improved methods for forming apattern on a substrate, such as method employing a patterned mask. SeeU.S. Pat. No. 4,735,890 to Nakane et al.; U.S. Pat. No. 5,147,763 toKamitakahara et al.; U.S. Pat. No. 5,259,926 to Kuwabara et al.; andInternational PCT Publication No. WO 99/54786 to Jackson et al., each ofwhich is incorporated herein by reference in their entirety.

There also is a need in the art for an improved method for formingisolated structures that can be considered “engineered” structures,including but not limited to particles, shapes, and parts. Usingtraditional IL methods, the scum layer that almost always forms betweenstructures acts to connect or link structures together, thereby makingit difficult, if not impossible to fabricate and/or harvest isolatedstructures.

There also is a need in the art for an improved method for formingmicro- and nanoscale charged particles, in particular polymer electrets.The term “polymer electrets” refers to dielectrics with stored charge,either on the surface or in the bulk, and dielectrics with orienteddipoles, frozen-in, ferrielectric, or ferroelectric. On the macro scale,such materials are used, for example, for electronic packaging andcharge electret devices, such as microphones and the like. See Kressman.R., et al., Space-Charge Electrets, Vol. 2, Laplacian Press, 1999; andHarrison, J. S., et al., Piezoelectic Polymers, NASA/CR-2001-211422,ICASE Report No. 200143. Poly(vinylidene fluoride) (PVDF) is one exampleof a polymer electret material. In addition to PVDF, charge electretmaterials, such as polypropylene (PP), Teflon-fluorinated ethylenepropylene (FEP), and polytetrafluoroethylene (PTFE), also are consideredpolymer electrets.

Further, there is a need in the art for improved methods for deliveringtherapeutic agents, such as drugs, non-viral gene vectors, DNA, RNA,RNAi, and viral particles, to a target. See Biomedical Polymers,Shalaby, S. W., ed., Harner/Gardner Publications, Inc., Cincinnati,Ohio, 1994; Polymeric Biomaterials, Dumitrin, S., ed., Marcel Dekkar,Inc., New York, N.Y., 1994; Park, K., et al., Biodegradable Hydrogelsfor Drug Delivery, Technomic Publishing Company, Inc., Lancaster, Pa.,1993; Gumargalieva, et al., Biodegradation and Biodeterioration ofPolymers: Kinetic Aspects, Nova Science Publishers, Inc., Commack, N.Y.,1998; Controlled Drug Delivery, American Chemical Society SymposiumSeries 752, Park, K., and Mrsny, R. J., eds., Washington, D.C., 2000;Cellular Drug Delivery: Principles and Practices, Lu, D. R., and Oie,S., eds., Humana Press, Totowa, N.J., 2004; and Bioreversible Carriersin Drug Design: Theory and Applications, Roche, E. B., ed., PergamonPress, New York, N.Y., 1987. For a description of representativetherapeutic agents for use in such delivery methods, see U.S. Pat. No.6,159,443 to Hallahan, which is incorporated herein by reference in itsentirety.

There is also a need in the art for an improved method for forming superabsorbent particles. These particles can be used for specialtypackaging, wire waterblocking, filtration, medical markets, spillcontrol, therapy packs, composites and laminates, water retention.

There is also a need in the art for improved methods to createpolymorphs. Polymorphs exist when there is more than one way for theparticles of a particular substance to arrange themselves into acrystalline array. Different polymorphs of the same substance can havevastly different physical and chemical properties. Invariably, one ofthe crystal forms may be more stable or easier to handle than anotheralthough the conditions under which the various crystal forms appearsmay be so close as to be very difficult to control on the large scale.This effect can create differences in the bioavailability of the drugwhich leads to inconsistencies in efficacy. See “Drug polymorphism anddosage form design: a practical perspective” Adv. Drug Deliv. Rev.,Singhal D, Curatolo W. Feb. 23, 2004; 56(3):33547; Generic Drug ProductDevelopment Solid Oral Dosage Forms, Shargel, L., ed., Marcel Dekker,New York, 2005.

In sum, there exists a need in the art to identify new materials for usein imprint lithographic techniques. More particularly, there is a needin the art for methods for the fabrication of structures at the hundredsof micron level down to sub-100 nm feature sizes. Additionally, there isa need in the art for improved methods for polymorph creation.

SUMMARY

In some embodiments, the present invention includes a particle having ageometric solid shape, wherein a maximum cross-sectional dimension ofthe particle is less than about 1 micrometer. In alternativeembodiments, the maximum cross-sectional dimension is between about 5nanometers and about 1 micrometer; between about 10 nanometers and about1 micrometer; less than about 800 nanometers; less than about 750nanometers; less than about 500 nanometers; less than about 300nanometers; less than about 250 nanometers; less than about 200nanometers; less than about 150 nanometers; or less than about 100nanometers. According to some embodiments, the present inventionincludes a plurality of substantially congruent particles. In someembodiments, the particles include a maximum cross-sectional dimensionas described herein.

According to some embodiments, the particle includes a reaction productof a methacrylate; a reaction product of an acrylate; a reaction productof an epoxy; a reaction product of a free radical polymerization; athermoplastic material; an organic material; an imaging agent; a drug; atreatment agent; an antibiotic; biologic material; a soluble material; abiodegradable material; a hydrophilic material; a hydrophobic material;an inorganic material; a polymer material; a small molecule; a ceramic;a metal; a material cured by applying actinic radiation, such as UVlight; a material that hardens through evaporation, such as throughevaporation of a solvent; a material that hardens through a chemicalreaction; or a material that hardens through a change in temperature,such as through a melt transition of the material or a transitioningfrom between a flowable and non-flowable configuration. In someembodiments, the particle has a modulus from about 0.1 MPa to about 500MPa. In other embodiments, the particle has a modulus of about 1 MPa toabout 100 MPa. In some embodiments, the particle includes a porogen.

In other embodiments, a particle of the present invention has anengineered geometric shape and a volume of the particle is less thanabout 4200 cubic micrometers. In some embodiments, a particle of thepresent invention has a geometric solid shape, wherein a maximumcross-sectional dimension of the particle is less than about 10micrometer and the particle includes a biologic material. In otherembodiments, a particle of the present invention has a geometric solidshape, wherein a maximum cross-sectional dimension of the particle isless than about 10 micrometer and the particle includes a drug.

In alternative embodiments of the present invention, a nanostructureincludes a layer of a first material and a structure having a geometricsolid shape configured from a second material, wherein the structure iscoupled with the layer and the structure has a maximum cross-sectionaldimension of less than about 10 micrometers. In alternative embodiments,the structure includes a maximum cross-sectional dimension between about5 nanometers and about 5 micrometers; between about 10 nanometers andabout 2 micrometers; between about 10 nanometers and about 1 micrometer;less than about 1 micrometer; less than about 750 nanometers; less thanabout 500 nanometers; less than about 300 nanometers; less than about250 nanometers; less than about 200 nanometers; less than about 150nanometers; or less than about 100 nanometers.

According to some embodiments, a plurality of substantially congruentstructures are coupled with the layer. In some embodiments, theplurality of structures are arranged in a substantially predeterminedorientation and in other embodiments, the plurality of structures arearranged in a substantially ordered array.

According to some embodiments, the second material of the nanostructureincludes a reaction product of a methacrylate; a reaction product of anacrylate; a reaction product of an epoxy; a reaction product of a freeradical polymerization; a thermoplastic material; an organic material;an imaging agent; a drug; a treatment agent; an antibiotic; biologicmaterial; a porogen; a soluble material; a biodegradable material; ahydrophilic material; a hydrophobic material; an inorganic material; apolymer material; a small molecule; a ceramic; a metal; a material curedby applying actinic radiation, such as UV light; a material that hardensthrough evaporation, such as through evaporation of a solvent; amaterial that hardens through a chemical reaction; or a material thathardens through a change in temperature, such as through a melttransition of the second material or a transitioning from between aflowable and non-flowable configuration. According to other embodiments,the first material and the second material include the same composition.

In some embodiments of the present invention, the layer of materialincludes a thickness of about twice a dimension of the structure. Inother embodiments, the layer of material includes a thickness of aboutequal a dimension of the structure. In still other embodiments, thelayer of material includes a thickness of about one half a dimension ofthe structure. According to some embodiments, structures of more thanone size are coupled with the layer. In some embodiments, structures ofmore than one shape are coupled with the layer. In other embodiments, anagent is configured to couple the structure to the layer by interactionssuch as covalent binding, ionic bonding, electrostatic binding, surfaceenergy, hydrogen bonding, van der Waals forces, other intra- andinter-molecular forces, adhesives, or a magnetic force.

The present invention also includes methods for fabricating particles,particles attached to a scum layer and particles adhered to films. Insome embodiments, a method of fabricating arrayed nanostructuresincludes placing a first material into a recess in a polymer mold wherethe recess is less than about 10 micrometers in a maximumcross-sectional dimension, hardening the first material to form aparticle having a geometric solid shape substantially corresponding tothe recess, removing the particle from the recess, and coupling theparticle with a film. In alternative embodiments, the maximumcross-sectional dimension is between about 5 nanometers and about 5micrometers; between about 10 nanometers and about 2 micrometers;between about 10 nanometers and about 1 micrometer; less than aboutmicrometer; less than about 750 nanometers; less than about 500nanometers; less than about 300 nanometers; less than about 250nanometers; less than about 200 nanometers; less than about 150nanometers; or less than about 100 nanometers. In some embodiments, aplurality of substantially congruent particles are fabricated by placingthe first material into a plurality of substantially congruent recessesin the polymer mold, wherein the recess is less than about 10micrometers in a maximum cross-sectional dimension and hardening thefirst material to form a plurality of substantially congruent particles,each having a geometric solid shape substantially corresponding to therecess in which it was hardened. Next, the particles are removed fromthe recesses and coupled with a film. In some embodiments, the polymermold includes a fluorinated polymer. In other embodiments, thefluorinated polymer includes a perfluoropolyether.

BRIEF DESCRIPTION OF THE DRAWINGS

Reference is made to the accompanying drawings in which are shownillustrative embodiments of the presently disclosed subject matter, fromwhich its novel features and advantages will be apparent.

FIGS. 1A-1D are a schematic representation of an embodiment of thepresently disclosed method for preparing a patterned template;

FIGS. 2A-2F are a schematic representation of the presently disclosedmethod for forming one or more micro- and/or nanoscale particles;

FIGS. 3A-3F are a schematic representation of the presently disclosedmethod for preparing one or more spherical particles;

FIGS. 4A-4D are a schematic representation of the presently disclosedmethod for fabricating charged polymeric particles. FIG. 4A representsthe electrostatic charging of the molded particle during polymerizationor crystallization; FIG. 4B represents a charged nano-disc; FIG. 4Crepresents typical random juxtapositioning of uncharged nano-discs; andFIG. 4D represents the spontaneous aggregation of charged nano-discsinto chain-like structures;

FIGS. 5A-5C are a schematic illustration of multilayer particles thatcan be formed using the presently disclosed soft lithography method;

FIGS. 6A-6C are a schematic representation of the presently disclosedmethod for making three-dimensional nanostructures using a softlithography technique;

FIGS. 7A-7F are a schematic representation of an embodiment of thepresently disclosed method for preparing a multi-dimensional complexstructure;

FIGS. 8A-8E are a schematic representation of the presently disclosedimprint lithography process resulting in a “scum layer”;

FIGS. 9A-9E are a schematic representation of the presently disclosedimprint lithography method, which eliminates the “scum layer” by using afunctionalized, non-wetting patterned template and a non-wettingsubstrate;

FIGS. 10A-10E are a schematic representation of the presently disclosedsolvent-assisted micro-molding (SAMIM) method for forming a pattern on asubstrate;

FIG. 11 is a scanning electron micrograph of a silicon master including3-μm arrow-shaped patterns;

FIG. 12 is a scanning electron micrograph of a silicon master including500 nm conical patterns that are <50 nm at the tip;

FIG. 13 is a scanning electron micrograph of a silicon master including200 nm trapezoidal patterns;

FIG. 14 is a scanning electron micrograph of 200-nm isolated trapezoidalparticles of poly(ethylene glycol) (PEG) diacrylate;

FIG. 15 is a scanning electron micrograph of 500-nm isolated conicalparticles of PEG diacrylate;

FIG. 16 is a scanning electron micrograph of 3-μm isolated arrow-shapedparticles of PEG diacrylate;

FIG. 17 is a scanning electron micrograph of 200-nm×750-nm×250-nmrectangular shaped particles of PEG diacrylate;

FIG. 18 is a scanning electron micrograph of 200-nm isolated trapezoidalparticles of trimethylolpropane triacrylate (TMPTA);

FIG. 19 is a scanning electron micrograph of 500-nm isolated conicalparticles of TMPTA;

FIG. 20 is a scanning electron micrograph of 500-nm isolated conicalparticles of TMPTA, which have been printed using an embodiment of thepresently described non-wetting imprint lithography method and harvestedmechanically using a doctor blade;

FIG. 21 is a scanning electron micrograph of 200-nm isolated trapezoidalparticles of poly(lactic acid) (PLA);

FIG. 22 is a scanning electron micrograph of 200-nm isolated trapezoidalparticles of poly(lactic acid) (PLA), which have been printed using anembodiment of the presently described non-wetting imprint lithographymethod and harvested mechanically using a doctor blade;

FIG. 23 is a scanning electron micrograph of 3-μm isolated arrow-shapedparticles of PLA;

FIG. 24 is a scanning electron micrograph of 500-nm isolatedconical-shaped particles of PLA;

FIG. 25 is a scanning electron micrograph of 200-nm isolated trapezoidalparticles of poly(pyrrole) (Ppy);

FIG. 26 is a scanning electron micrograph of 3-μm arrow-shaped Ppyparticles;

FIG. 27 is a scanning electron micrograph of 500-nm conical shaped Ppyparticles;

FIGS. 28A-28C are fluorescence confocal micrographs of 200-nm isolatedtrapezoidal particles of PEG diacrylate that contain fluorescentlytagged DNA. FIG. 28A is a fluorescent confocal micrograph of 200 nmtrapezoidal PEG nanoparticles which contain 24-mer DNA strands that aretagged with CY-3. FIG. 28B is optical micrograph of the 200-nm isolatedtrapezoidal particles of PEG diacrylate that contain fluorescentlytagged DNA. FIG. 28C is the overlay of the images provided in FIGS. 28Aand 28B, showing that every particle contains DNA;

FIG. 29 is a scanning electron micrograph of fabrication of 200-nmPEG-diacrylate nanoparticles using “double stamping”;

FIG. 30 is an atomic force micrograph image of 140-nm lines of TMPTAseparated by distance of 70 nm that were fabricated using a PFPE mold;

FIGS. 31A and 31B are a scanning electron micrograph of mold fabricationfrom electron-beam lithographically generated masters. FIG. 31A is ascanning electron micrograph of silicon/silicon oxide masters of 3micron arrows. FIG. 31B is a scanning electron micrograph ofsilicon/silicon oxide masters of 200-nm×800-nm bars;

FIGS. 32A and 32B are an optical micrographic image of mold fabricationfrom photoresist masters. FIG. 32A is a SU-8 master. FIG. 32B is aPFPE-DMA mold templated from a photolithographic master;

FIGS. 33A and 33B are an atomic force micrograph of mold fabricationfrom Tobacco Mosaic Virus templates. FIG. 33A is a master. FIG. 33B is aPFPE-DMA mold templated from a virus master;

FIGS. 34A and 34B are an atomic force micrograph of mold fabricationfrom block copolymer micelle masters. FIG. 34A is apolystyrene-polyisoprene block copolymer micelle. FIG. 34B is a PFPE-DMAmold templated from a micelle master;

FIGS. 35A and 35B are an atomic force micrograph of mold fabricationfrom brush polymer masters. FIG. 35A is a brush polymer master. FIG. 35Bis a PFPE-DMA mold templated from a brush polymer master;

FIGS. 36A-36D are schematic representations of one embodiment of amethod for functionalizing particles of the presently disclosed subjectmatter;

FIGS. 37A-37F are schematic representations of one embodiment of amethod of the presently disclosed subject matter for harvestingparticles from an article;

FIGS. 38A-38G are schematic representations of one embodiment of amethod of the presently disclosed subject matter for harvestingparticles from an article;

FIGS. 39A-39F are schematic representations of one embodiment of oneprocess of the presently disclosed subject matter for imprintlithography wherein 3-dimensional features are patterned;

FIGS. 40A-40D schematic representations of one embodiment of one processof the presently disclosed subject matter for harvesting particles froman article;

FIGS. 41A-41E show a sequence of forming small particles throughevaporation according to an embodiment of the presently disclosedsubject matter;

FIG. 42 shows doxorubicin containing particles after removal from atemplate according to an embodiment of the presently disclosed subjectmatter;

FIG. 43 shows a structure patterned with nano-cylindrical shapesaccording to an embodiment of the presently disclosed subject matter;

FIG. 44 shows a sequence of molecular imprinting according to anembodiment of the presently disclosed subject matter;

FIG. 45 shows a labeled particle associated with a cell according to anembodiment of the presently disclosed subject matter;

FIG. 46 shows a labeled particle associated with a cell according to anembodiment of the presently disclosed subject matter;

FIG. 47 shows particles fabricated through an open molding techniqueaccording to some embodiments of the present invention;

FIG. 48 shows a process for coating a seed and seeds coated from theprocess according to some embodiments of the present invention;

FIG. 49A shows a silicon template having a 2-dimensional array of 200 nmtrapezoid recesses;

FIG. 49B shows 200 nm trapezoidal PLA particles fabricated according toan embodiment of the present invention;

FIG. 49C shows 200 nm trapezoidal poly(pyrrole) (PPy) particlesfabricated according to an embodiment of the present invention;

FIG. 49D shows 200 nm trapezoidal trimethylopropane triacrylate (TMPTA)particles fabricated according to an embodiment of the presentinvention;

FIGS. 50A-50F show PEG particles of different shapes and sizesfabricated according to embodiments of the present invention;

FIG. 51 shows a DLS trace showing congruent particles fabricatedaccording to embodiments of the present invention;

FIG. 52A shows discrete 200 nm PEG particles fabricated according toembodiments of the present invention;

FIG. 52B shows 200 nm PEG particles connected by a PEG film afterdragging a blade across a surface to roll up the film according toembodiments of the present invention;

FIG. 52C shows 200 nm PEG particles coupled with a glass slide with acyanoacrylate monomer according to embodiments of the present invention;and

FIGS. 53A and 53B show 2 micrometer TMPTA particles having Bosch-typeetch lines on sidewalls of the particles according to embodiments of thepresent invention.

DETAILED DESCRIPTION

The presently disclosed subject matter will now be described more fullyhereinafter with reference to the accompanying Examples, in whichrepresentative embodiments are shown. The presently disclosed subjectmatter can, however, be embodied in different forms and should not beconstrued as limited to the embodiments set forth herein. Rather, theseembodiments are provided so that this disclosure will be thorough andcomplete, and will fully convey the scope of the embodiments to thoseskilled in the art.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this presently described subject matter belongs. Allpublications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety.

Throughout the specification and claims, a given chemical formula orname shall encompass all optical and stereoisomers, as well as racemicmixtures where such isomers and mixtures exist.

I. Materials

The presently disclosed subject matter broadly describes solventresistant, low surface energy polymeric materials, derived from castinglow viscosity liquid materials onto a master template and then curingthe low viscosity liquid materials to generate a patterned template foruse in high-resolution soft or imprint lithographic applications, suchas micro- and nanoscale replica molding. In some embodiments, thepatterned template includes a solvent resistant, elastomer-basedmaterial, such as but not limited to a fluorinated elastomer-basedmaterial.

Further, the presently disclosed subject matter describes the firstnano-contact molding of organic materials to generate high fidelityfeatures using an elastomeric mold. Accordingly, the presently disclosedsubject matter describes a method for producing free-standing, isolatedmicro- and nanostructures of any shape using soft or imprint lithographytechniques. Representative micro- and nanostructures include but are notlimited to micro- and nanoparticles, and micro- and nano-patternedsubstrates.

The nanostructures described by the presently disclosed subject mattercan be used in several applications, including, but not limited to,semiconductor manufacturing, such as molding etch barriers without scumlayers for the fabrication of semiconductor devices; crystals; materialsfor displays; photovoltaics; a solar cell device; optoelectronicdevices; routers; gratings; radio frequency identification (RFID)devices; catalysts; fillers and additives; detoxifying agents; etchbarriers; atomic force microscope (AFM) tips; parts for nano-machines;the delivery of a therapeutic agent, such as a drug or genetic material;cosmetics; chemical mechanical planarization (CMP) particles; and porousparticles and shapes of any kind that will enable the nanotechnologyindustry.

Representative solvent resistant elastomer-based materials include butare not limited to fluorinated elastomer-based materials. As usedherein, the term “solvent resistant” refers to a material, such as anelastomeric material that neither swells nor dissolves in commonhydrocarbon-based organic solvents or acidic or basic aqueous solutions.Representative fluorinated elastomer-based materials include but are notlimited to perfluoropolyether (PFPE)-based materials. A photocurableliquid PFPE exhibits desirable properties for soft lithography. Arepresentative scheme for the synthesis and photocuring of functionalPFPEs is provided in Scheme 1.

According to another embodiment, a material according to the presentlydisclosed subject matter includes one or more of a photo-curableconstituent, a thermal-curable constituent, and mixtures thereof. In oneembodiment, the photo-curable constituent is independent from thethermal-curable constituent such that the material can undergo multiplecures. A material having the ability to undergo multiple cures isuseful, for example, in forming layered devices. For example, a liquidmaterial having photo-curable and thermal-curable constituents canundergo a first cure to form a first device through, for example, aphotocuring process or a thermal curing process. Then the photocured orthermal cured first device can be adhered to a second device of the samematerial or any material similar thereto that will thermally cure orphotocure and bind to the material of the first device. By positioningthe first device and second device adjacent one another and subjectingthe first and second devices to a thermalcuring or photocuring process,whichever component that was not activated on the first curing can becured by a subsequent curing step. Thereafter, either the thermalcureconstituents of the first device that was left un-activated by thephotocuring process or the photocure constituents of the first devicethat were left un-activated by the first thermal curing, will beactivated and bind the second device. Thereby, the first and seconddevices become adhered together. It will be appreciated by one ofordinary skill in the art that the order of curing processes isindependent and a thermal-curing could occur first followed by aphotocuring or a photocuring could occur first followed by a thermalcuring.

According to yet another embodiment, multiple thermo-curableconstituents can be included in the material such that the material canbe subjected to multiple independent thermal-cures. For example, themultiple thermo-curable constituents can have different activationtemperature ranges such that the material can undergo a firstthermal-cure at a first temperature range and a second thermal-cure at asecond temperature range.

Additional schemes for the synthesis of functional perfluoropolyethersare provided in Examples 7.1 through 7.6.

According to one embodiment this PFPE material has a surface energybelow about 30 mN/m. According to another embodiment the surface energyof the PFPE is between about 10 mN/m and about 20 mN/m. According to aanother embodiment, the PFPE has a low surface energy of between about12 mN/m and about 15 mN/m. The PFPE is non-toxic, UV transparent, andhighly gas permeable; and cures into a tough, durable, highlyfluorinated elastomer with excellent release properties and resistanceto swelling. The properties of these materials can be tuned over a widerange through the judicious choice of additives, fillers, reactiveco-monomers, and functionalization agents. Such properties that aredesirable to modify, include, but are not limited to, modulus, tearstrength, surface energy, permeability, functionality, mode of cure,solubility and swelling characteristics, and the like. The non-swellingnature and easy release properties of the presently disclosed PFPEmaterials allows for nanostructures to be fabricated from any material.Further, the presently disclosed subject matter can be expanded to largescale rollers or conveyor belt technology or rapid stamping that allowfor the fabrication of nanostructures on an industrial scale.

In some embodiments, the patterned template includes a solventresistant, low surface energy polymeric material derived from castinglow viscosity liquid materials onto a master template and then curingthe low viscosity liquid materials to generate a patterned template. Insome embodiments, the patterned template includes a solvent resistantelastomeric material.

In some embodiments, at least one of the patterned template andsubstrate includes a material selected from the group including aperfluoropolyether material, a fluoroolefin material, an acrylatematerial, a silicone material, a styrenic material, a fluorinatedthermoplastic elastomer (TPE), a triazine fluoropolymer, aperfluorocyclobutyl material, a fluorinated epoxy resin, and afluorinated monomer or fluorinated oligomer that can be polymerized orcrosslinked by a metathesis polymerization reaction.

In some embodiments, the perfluoropolyether material includes a backbonestructure selected from the group including:

wherein X is present or absent, and when present includes an endcappinggroup.

In some embodiments, the fluoroolefin material is selected from thegroup including:

wherein CSM includes a cure site monomer.

In some embodiments, the fluoroolefin material is made from monomerswhich include tetrafluoroethylene, vinylidene fluoride,hexafluoropropylene, 2,2-bis(trifluoromethyl)-4,5-difluoro-1,3-dioxole,a functional fluoroolefin, functional acrylic monomer, and a functionalmethacrylic monomer.

In some embodiments, the silicone material includes a fluoroalkylfunctionalized polydimethylsiloxane (PDMS) having the followingstructure:

wherein:

R is selected from the group including an acrylate, a methacrylate, anda vinyl group; and

Rf includes a fluoroalkyl chain.

In some embodiments, the styrenic material includes a fluorinatedstyrene monomer selected from the group including:

wherein Rf includes a fluoroalkyl chain.

In some embodiments, the acrylate material includes a fluorinatedacrylate or a fluorinated methacrylate having the following structure:

wherein:

R is selected from the group including H, alkyl, substituted alkyl,aryl, and substituted aryl; and

Rf includes a fluoroalkyl chain.

In some embodiments, the triazine fluoropolymer includes a fluorinatedmonomer. In some embodiments, the fluorinated monomer or fluorinatedoligomer that can be polymerized or crosslinked by a metathesispolymerization reaction includes a functionalized olefin. In someembodiments, the functionalized olefin includes a functionalized cyclicolefin.

In some embodiments, at least one of the patterned template and thesubstrate has a surface energy lower than about 18 mN/m. In someembodiments, at least one of the patterned template and the substratehas a surface energy lower than about 15 mN/m. According to a furtherembodiment the patterned template and/or the substrate has a surfaceenergy between about 10 mN/m and about 20 mN/m. According to anotherembodiment, the patterned template and/or the substrate has a lowsurface energy of between about 12 mN/m and about 15 mN/m.

From a property point of view, the exact properties of these moldingmaterials can be adjusted by adjusting the composition of theingredients used to make the materials. In particular the modulus can beadjusted from low (approximately 1 MPa) to multiple GPa.

II. Formation of Isolated Micro- and/or Nanoparticles

In some embodiments, the presently disclosed subject matter provides amethod for making isolated micro- and/or nanoparticles. In someembodiments, the process includes initially forming a patternedsubstrate. Turning now to FIG. 1A, a patterned master 100 is provided.Patterned master 100 includes a plurality of non-recessed surface areas102 and a plurality of recesses 104. In some embodiments, patternedmaster 100 includes an etched substrate, such as a silicon wafer, whichis etched in the desired pattern to form patterned master 100.

Referring now to FIG. 1B, a liquid material 106, for example, a liquidfluoropolymer composition, such as a PFPE-based precursor, is thenpoured onto patterned master 100. Liquid material 106 is treated bytreating process T_(r), for example exposure to UV light, actinicradiation, or the like, thereby forming a treated liquid material 108 inthe desired pattern.

Referring now to FIGS. 1C and 1D, a force F_(r) is applied to treatedliquid material 108 to remove it from patterned master 100. As shown inFIGS. 1C and 1D, treated liquid material 108 includes a plurality ofrecesses 110, which are mirror images of the plurality of non-recessedsurface areas 102 of patterned master 100. Continuing with FIGS. 1C and1D, treated liquid material 108 includes a plurality of first patternedsurface areas 112, which are mirror images of the plurality of recesses104 of patterned master 100. Treated liquid material 108 can now be usedas a patterned template for soft lithography and imprint lithographyapplications. Accordingly, treated liquid material 108 can be used as apatterned template for the formation of isolated micro- andnanoparticles. For the purposes of FIGS. 1A-1D, 2A-2E, and 3A-3F, thenumbering scheme for like structures is retained throughout, wherepossible.

Referring now to FIG. 2A, in some embodiments, a substrate 200, forexample, a silicon wafer, is treated or is coated with a non-wettingmaterial 202. In some embodiments, non-wetting material 202 includes anelastomer (such a solvent resistant elastomer, including but not limitedto a PFPE elastomer) that can be further exposed to UV light and curedto form a thin, non-wetting layer on the surface of substrate 200.Substrate 200 also can be made non-wetting by treating substrate 200with non-wetting agent 202, for example a small molecule, such as analkyl- or fluoroalkyl-silane, or other surface treatment. Continuingwith FIG. 2A, a droplet 204 of a curable resin, a monomer, or a solutionfrom which the desired particles will be formed is then placed on thecoated substrate 200.

Referring now to FIG. 2A and FIG. 2B, patterned template 108 (as shownin FIG. 1D) is then contacted with droplet 204 of a particle precursormaterial so that droplet 204 fills the plurality of recessed areas 110of patterned template 108.

Referring now to FIGS. 2C and 2D, a force F_(a) is applied to patternedtemplate 108. While not wishing to be bound by any particular theory,once force F_(a) is applied, the affinity of patterned template 108 fornon-wetting coating or surface treatment 202 on substrate 200 incombination with the non-wetting behavior of patterned template 108 andsurface treated or coated substrate 200 causes droplet 204 to beexcluded from all areas except for recessed areas 110. Further, inembodiments essentially free of non-wetting or low wetting material 202with which to sandwich droplet 204, a “scum” layer forms thatinterconnects the objects being stamped.

Continuing with FIGS. 2C and 2D, the particle precursor material fillingrecessed areas 110, e.g., a resin, monomer, solvent, combinationsthereof, or the like, is then treated by a treating process T_(r), e.g.,photocured, UV-light treated, or actinic radiation treated, throughpatterned template 108 or thermally cured while under pressure, to forma plurality of micro- and/or nanoparticles 206. In some embodiments, amaterial, including but not limited to a polymer, an organic compound,or an inorganic compound, can be dissolved in a solvent, patterned usingpatterned template 108, and the solvent can be released.

Continuing with FIGS. 2C and 2D, once the material filling recessedareas 110 is treated, patterned template 108 is removed from substrate200. Micro- and/or nanoparticles 206 are confined to recessed areas 110of patterned template 108. In some embodiments, micro- and/ornanoparticles 206 can be retained on substrate 200 in defined regionsonce patterned template 108 is removed. This embodiment can be used inthe manufacture of semiconductor devices where essentially scum-layerfree features could be used as etch barriers or as conductive,semiconductive, or dielectric layers directly, mitigating or reducingthe need to use traditional and expensive photolithographic processes.

Referring now to FIGS. 2D and 2E, micro- and/or nanoparticles 206 can beremoved from patterned template 108 to provide freestanding particles bya variety of methods, which include but are not limited to: (1) applyingpatterned template 108 to a surface that has an affinity for theparticles 206; (2) deforming patterned template 108, or using othermechanical methods, including sonication, in such a manner that theparticles 206 are naturally released from patterned template 108; (3)swelling patterned template 108 reversibly with supercritical carbondioxide or another solvent that will extrude the particles 206; (4)washing patterned template 108 with a solvent that has an affinity forthe particles 206 and will wash them out of patterned template 108; (5)applying patterned template 108 to a liquid that when hardenedphysically entraps particles 206; (6) applying patterned template 108 toa material that when hardened has a chemical and/or physical interactionwith particles 206.

In some embodiments, the method of producing and harvesting particlesincludes a batch process. In some embodiments, the batch process isselected from one of a semi-batch process and a continuous batchprocess. Referring now to FIG. 2F, an embodiment of the presentlydisclosed subject matter wherein particles 206 are produced in acontinuous process is schematically presented. An apparatus 199 isprovided for carrying out the process. Indeed, while FIG. 2Fschematically presents a continuous process for particles, apparatus 199can be adapted for batch processes, and for providing a pattern on asubstrate continuously or in batch, in accordance with the presentlydisclosed subject matter and based on a review of the presentlydisclosed subject matter by one of ordinary skill in the art.

Continuing, then, with FIG. 2F, droplet 204 of liquid material isapplied to substrate 200′ via reservoir 203. Substrate 200′ can becoated or not coated with a non-wetting agent. Substrate 200′ andpattern template 108′ are placed in a spaced relationship with respectto each other and are also operably disposed with respect to each otherto provide for the conveyance of droplet 204 between patterned template108′ and substrate 200′. Conveyance is facilitated through the provisionof pulleys 208, which are in operative communication with controller201. By way of representative non-limiting examples, controller 201 caninclude a computing system, appropriate software, a power source, aradiation source, and/or other suitable devices for controlling thefunctions of apparatus 199. Thus, controller 201 provides for power forand other control of the operation of pulleys 208 to provide for theconveyance of droplet 204 between patterned template 108′ and substrate200′. Particles 206 are formed and treated between substrate 200′ andpatterned template 108′ by a treating process T_(R), which is alsocontrolled by controller 201. Particles 206 are collected in aninspecting device 210, which is also controlled by controller 201.Inspecting device 210 provides for one of inspecting, measuring, andboth inspecting and measuring one or more characteristics of particles206. Representative examples of inspecting devices 210 are disclosedelsewhere herein.

By way of further exemplifying embodiments of particle harvestingmethods described herein, reference is made to FIGS. 37A-37F and FIGS.38A-38G. In FIGS. 37A-37C and FIGS. 38A-38C particles which are producedin accordance with embodiments described herein remain in contact withan article 3700, 3800 having an affinity for particles 3705 and 3805respectively. In one embodiment, article 3700 is a patterned template ormold as described herein. In one embodiment, article 3800 is a substrateas described herein.

Referring now to FIGS. 37D-37F and FIGS. 38D-38G, material 3720, 3820having an affinity for particles 3705, 3805 is put into contact withparticles 3705, 3805 while particles 3705, 3805 remain in connectionwith articles 3700, 3800. In the embodiment of FIG. 37D, material 3720is disposed on surface 3710. In the embodiment of FIG. 38D, material3820 is applied directly to article 3800 having particles 3820. Asillustrated in FIGS. 37E, 38D in some embodiments, article 3700, 3800 isput in engaging contact with material 3720, 3820. In one embodimentmaterial 3720, 3820 is thereby dispersed to coat at least a portion ofsubstantially all of particles 3705, 3805 while particles 3705, 3805 areattached to article 3700, 3800 (e.g., a patterned template). In oneembodiment, illustrated in FIGS. 37F and 38F, articles 3700, 3800 aresubstantially disassociated with material 3720, 3820. In one embodiment,material 3720, 3820 has a higher affinity for particles 3705, 3805 thanthe affinity between article 3700, 3800 and particles 3705, 3805. InFIGS. 37F and 38F, the disassociation of article 3700, 3800 frommaterial 3720, 3820 thereby releases particles 3705, 3805 from article3700, 3800 leaving particles 3705, 3805 attached to material 3720, 3820.

In one embodiment material 3720, 3820 has an affinity for particles 3705and 3805. For example, in some embodiments, material 3720, 3820 includesan adhesive or sticky surface when applied to article 3700, 3800. Inother embodiments, material 3720, 3820 undergoes a transformation afterit is brought into contact with article 3700, 3800. In some embodimentsthat transformation is an inherent characteristic of material 3705,3805. In other embodiments, material 3705, 3805 is treated to induce thetransformation. For example, in one embodiment material 3720, 3820 is anepoxy that hardens after it is brought into contact with article 3700,3800. Thus when article 3700, 3800 is pealed away from the hardenedepoxy, particles 3705, 3805 remain engaged with the epoxy and notarticle 3700, 3800. In other embodiments, material 3720, 3820 is waterthat is cooled to form ice. Thus, when article 3700, 3800 is strippedfrom the ice, particles 3705, 3805 remain in communication with the iceand not article 3700, 3800. In one embodiment, the particle-containingice can be melted to create a liquid with a concentration of particles3705, 3805. In some embodiments, material 3705, 3805 include, withoutlimitation, one or more of a carbohydrate, an epoxy, a wax, polyvinylalcohol, polyvinyl pyrrolidone, polybutyl acrylate, a polycyano acrylateand polymethyl methacrylate. In some embodiments, material 3720, 3820includes, without limitation, one or more of liquids, solutions,powders, granulated materials, semi-solid materials, suspensions,combinations thereof, or the like.

Thus, in some embodiments, the method for forming and harvesting one ormore particles includes:

-   -   (a) providing a patterned template and a substrate, wherein the        patterned template includes a first patterned template surface        having a plurality of recessed areas formed therein;    -   (b) disposing a volume of liquid material in or on at least one        of:        -   (i) the first patterned template surface;        -   (ii) the plurality of recessed areas; and/or        -   (iii) a substrate; and    -   (c) forming one or more particles by one of:        -   (i) contacting the patterned template surface with the            substrate and treating the liquid material; and        -   (ii) treating the liquid material.

In some embodiments, the plurality of recessed areas includes aplurality of cavities. In some embodiments, the plurality of cavitiesincludes a plurality of structural features. In some embodiments, theplurality of structural features have a dimension ranging from about 10microns to about 1 nanometer in size. In some embodiments, the pluralityof structural features have a dimension ranging from about 1 micron toabout 100 nm in size. In some embodiments, the plurality of structuralfeatures have a dimension ranging from about 100 nm to about 1 nm insize. In some embodiments, the plurality of structural features have adimension in both the horizontal and vertical plane.

According to yet another embodiment the particles are harvested on afast dissolving substrate, sheet, or films. The film-forming agents caninclude, but are not limited to pullulan, hydroxypropylmethyl cellulose,hydroxyethyl cellulose, hydroxypropyl cellulose, polyvinyl pyrrolidone,carboxymethyl cellulose, polyvinyl alcohol, sodium alginate,polyethylene glycol, xanthan gum, tragacanth gum, guar gum, acacia gum,arabic gum, polyacrylic acid, methylmethacrylate copolymer, carboxyvinylpolymer, amylose, high amylose starch, hydroxypropylated high amylosestarch, dextrin, pectin, chitin, chitosan, levan, elsinan, collagen,gelatin, zein, gluten, soy protein isolate, whey protein isolate,casein, combinations thereof, and the like. In some embodiments,pullulan is used as the primary filler. In still other embodiments,pullulan is included in amounts ranging from about 0.01 to about 99 wt%, preferably about 30 to about 80 wt %, more preferably from about 45to about 70 wt %, and even more preferably from about 60 to about 65 wt% of the film.

The film can further include water, plasticizing agents, natural and/orartificial flavoring agents, sulfur precipitating agents, salivastimulating agents, cooling agents, surfactants, stabilizing agents,emulsifying agents, thickening agents, binding agents, coloring agents,sweeteners, fragrances, combinations thereof, and the like.

Suitable sweeteners include both natural and artificial sweeteners.Examples of some sweeteners that can be used with the sheets of thepresently disclosed subject matter include, but are not limited to: (a)water-soluble sweetening agents, such as monosaccharides, disaccharidesand polysaccharides such as xylose, ribose, glucose (dextrose), mannose,galactose, fructose (levulose), sucrose (sugar), maltose, invert sugar(a mixture of fructose and glucose derived from sucrose), partiallyhydrolyzed starch, corn syrup solids, dihydrochalcones, monellin,steviosides, and glycyrrhizin; (b) water-soluble artificial sweeteners,such as the soluble saccharin salts, sodium or calcium saccharin salts,cyclamate salts, the sodium, ammonium or calcium salt of3,4-dihydro-6-methyl-1,2,3-oxathiazine-4-one-2, 2-dioxide, the potassiumsalt of 3,4-dihydro-6-methyl-1,2,3-oxathiazine4-one-2,2-dioxide(acesulfame-K), the free acid form of saccharin, and the like; (c)dipeptide based sweeteners, such as L-aspartic acid derived sweeteners,L-aspartyl-L-phenylalanine methyl ester (aspartame) and materialsdescribed in U.S. Pat. No. 3,492,131, which is incorporated herein byreference in its entirety,L-alpha-aspartyl-N-(2,2,4,4-tetramethyl-3-thietanyl)-D-alaninamidehydrate, methyl esters of L-aspartyl-L-phenylglycerin andL-aspartyl-L-2,5,dihydrophenyl-glycine,L-aspartyl-2,5-dihydro-L-phenylalanine,L-aspartyl-L-(1-cyclohexyen)-alanine, and the like; (d) water-solublesweeteners derived from naturally occurring water-soluble sweeteners,such as a chlorinated derivative of ordinary sugar (sucrose); and (e)protein based sweeteners, such as thaumatoccous danielli (Thaumatin Iand II) and the like.

In general, an effective amount of auxiliary sweetener is utilized toprovide the level of sweetness desired for a particular composition, andthis amount will vary with the sweetener selected. The amount willnormally be between about 0.01% to about 10% by weight of thecomposition when using an easily extractable sweetener. Thewater-soluble sweeteners described in category (a) above, are usuallyused in amounts of between about 0.01 to about 10 wt %, and preferablyin amounts of between about 2 to about 5 wt %. The sweeteners describedin categories (b)-(e) are generally used in amounts of between about0.01 to about 10 wt %, with between about 2 to about 8 wt % beingpreferred and between about 3 to about 6 wt % being most preferred.These amounts can be used to achieve a desired level of sweetnessindependent from the flavor level achieved from any optional flavor oilsused. Of course, sweeteners need not be added to films intended fornon-oral administration.

The flavorings that can be used in the films include natural andartificial flavors. These flavorings can be chosen from synthetic flavoroils and flavoring aromatics, and/or oils, oleo resins and extractsderived from plants, leaves, flowers, fruits, combinations thereof, andthe like. Representative flavor oils include: spearmint oil, cinnamonoil, peppermint oil, clove oil, bay oil, thyme oil, cedar leaf oil, oilof nutmeg, oil of sage, and oil of bitter almonds. Also useful areartificial, natural or synthetic fruit flavors, such as vanilla,chocolate, coffee, cocoa and citrus oil, including lemon, orange, grape,lime and grapefruit, and fruit essences including apple, pear, peach,strawberry, raspberry, cherry, plum, pineapple, apricot and so forth.These flavorings can be used individually or in admixture. Flavoringssuch as aldehydes and esters including cinnamyl acetate, cinnamaldehyde,citral, diethylacetal, dihydrocarvyl acetate, eugenyl formate,p-methylanisole, and so forth also can be used. Generally, any flavoringor food additive can be used, such as those described in Chemicals Usedin Food Processing, publication 1274 by the National Academy ofSciences, pages 63-258, which is incorporated herein by reference in itsentirety. Further examples of aldehyde flavorings include, but are notlimited to, acetaldehyde (apple); benzaldehyde (cherry, almond);cinnamic aldehyde (cinnamon); citral, i.e., alpha citral (lemon, lime);neral, i.e. beta citral (lemon, lime); decanal (orange, lemon); ethylvanillin (vanilla, cream); heliotropine, i.e., piperonal (vanilla,cream); vanillin (vanilla, cream); alpha-amyl cinnamaldehyde (spicyfruity flavors); butyraldehyde (butter, cheese); valeraldehyde (butter,cheese); citronellal; decanal (citrus fruits); aldehyde C-8 (citrusfruits); aldehyde C-9 (citrus fruits); aldehyde C-12 (citrus fruits);2-ethyl butyraldehyde (berry fruits); hexenal, i.e. trans-2 (berryfruits); tolyl aldehyde (cherry, almond); veratraldehyde (vanilla);2,6-dimethyl-5-heptenal, i.e. melonal (melon); 2-6-dimethyloctanal(green fruit); 2-dodecenal (citrus, mandarin); cherry; grape; mixturesthereof; and the like.

The amount of flavoring employed is normally a matter of preferencesubject to such factors as flavor type, individual flavor, strengthdesired, strength necessary to mask other less desirable flavors, andthe like. Thus, the amount can be varied to obtain the result desired inthe final product. In general, amounts of between about 0.1 to about 30wt % are useable with amounts of about 2 to about 25 wt % beingpreferred and amounts from about 8 to about 10 wt % are more preferred.

The films also can contain coloring agents or colorants. The coloringagents are used in amounts effective to produce a desired color. Thecoloring agents useful in the presently disclosed subject matter,include pigments, such as titanium dioxide, which can be incorporated inamounts of up to about 5 wt %, and preferably less than about 1 wt %.Colorants can also include natural food colors and dyes suitable forfood, drug and cosmetic applications. These colorants are known as FD&Cdyes and lakes. The materials acceptable for the foregoing spectrum ofuse are preferably water-soluble, and include FD&C Blue No. 2, which isthe disodium salt of 5,5-indigotindisulfonic acid. Similarly, the dyeknown as Green No. 3 comprises a triphenylmethane dye and is themonosodium salt of4-[4-N-ethyl-p-sulfobenzylamino)diphenyl-methylene]-[1-N-ethyl-N-p-sulfoniumbenzyl)-2,5-cyclo-hexadienimine]. A full recitation of all FD&C and D&Cdyes and their corresponding chemical structures can be found in theKirk-Othmer Encyclopedia of Chemical Technology, Volume 5, Pages857-884, which is incorporated herein by reference in its entirety.Furthermore, the materials and methods described in U.S. Pat. No.6,923,981 and the references cited therein, all of which areincorporated herein by reference, disclose appropriate fast-dissolvefilms for use with the particles of the presently disclosed subjectmatter.

After the particles are harvested on such sugar sheets, for example, thefast dissolving sheet can act as the delivery device. According to suchembodiments, the fast dissolve films can be placed on biological tissuesand as the film is dissolved and/or absorbed, the particles containedtherein are also dissolved or absorbed. The films can be configured fortransdermal delivery, trans mucosal delivery, nasal delivery, analdelivery, vaginal delivery, combinations thereof, and the like.

In some embodiments of the method for forming one or more particles, thepatterned template includes a solvent resistant, low surface energypolymeric material derived from casting low viscosity liquid materialsonto a master template and then curing the low viscosity liquidmaterials to generate a patterned template. In some embodiments, thepatterned template includes a solvent resistant elastomeric material.

In some embodiments, at least one of the patterned template andsubstrate includes a material selected from the group including aperfluoropolyether material, a fluoroolefin material, an acrylatematerial, a silicone material, a styrenic material, a fluorinatedthermoplastic elastomer (TPE), a triazine fluoropolymer, aperfluorocyclobutyl material, a fluorinated epoxy resin, and afluorinated monomer or fluorinated oligomer that can be polymerized orcrosslinked by a metathesis polymerization reaction.

In some embodiments, the perfluoropolyether material includes a backbonestructure selected from the group including:

wherein X is present or absent, and when present includes an endcappinggroup.

In some embodiments, the fluoroolefin material is selected from thegroup including:

wherein CSM includes a cure site monomer.

In some embodiments, the fluoroolefin material is made from monomerswhich include tetrafluoroethylene, vinylidene fluoride,hexafluoropropylene, 2,2-bis(trifluoromethyl)-4,5-difluoro-1,3-dioxole,a functional fluoroolefin, functional acrylic monomer, and a functionalmethacrylic monomer.

In some embodiments, the silicone material includes a fluoroalkylfunctionalized polydimethylsiloxane (PDMS) having the followingstructure:

wherein:

R is selected from the group including an acrylate, a methacrylate, anda vinyl group; and

Rf includes a fluoroalkyl chain.

In some embodiments, the styrenic material includes a fluorinatedstyrene monomer selected from the group including:

wherein Rf includes a fluoroalkyl chain.

In some embodiments, the acrylate material includes a fluorinatedacrylate or a fluorinated methacrylate having the following structure:

wherein:

R is selected from the group including H, alkyl, substituted alkyl,aryl, and substituted aryl; and

Rf includes a fluoroalkyl chain.

In some embodiments, the triazine fluoropolymer includes a fluorinatedmonomer. In some embodiments, the fluorinated monomer or fluorinatedoligomer that can be polymerized or crosslinked by a metathesispolymerization reaction includes a functionalized olefin. In someembodiments, the functionalized olefin includes a functionalized cyclicolefin.

In some embodiments, at least one of the patterned template and thesubstrate has a surface energy lower than 18 mN/m. In some embodiments,at least one of the patterned template and the substrate has a surfaceenergy lower than 15 mN/m. According to a further embodiment thepatterned template and/or the substrate has a surface energy betweenabout 10 mN/m and about 20 mN/m. According to another, the patternedtemplate and/or the substrate has a low surface energy of between about12 mN/m and about 15 mN/m.

In some embodiments, the substrate is selected from the group includinga polymer material, an inorganic material, a silicon material, a quartzmaterial, a glass material, and surface treated variants thereof. Insome embodiments, the substrate includes a patterned area.

According to an alternative embodiment, the PFPE material includes aurethane block as described and shown in the following structures:

According to an embodiment of the presently disclosed subject matter,PFPE urethane tetrafunctional methacrylate materials, such as the abovedescribed material, can be used as the materials and methods of thepresently disclosed subject matter or can be used in combination withother materials and methods described herein, as will be appreciated byone of ordinary skill in the art.

In some embodiments, the patterned template includes a patternedtemplate formed by a replica molding process. In some embodiments, thereplica molding process includes: providing a master template;contacting a liquid material with the master template; and curing theliquid material to form a patterned template.

In some embodiments, the master template includes, without limitation,one or more of a template formed from a lithography process, a naturallyoccurring template, combinations thereof, or the like. In someembodiments, the natural template is selected from one of a biologicalstructure and a self-assembled structure. In some embodiments, the oneof a biological structure and a self-assembled structure is selectedfrom the group including a naturally occurring crystal, an enzyme, avirus, a protein, a micelle, and a tissue surface.

In some embodiments, the method includes modifying the patternedtemplate surface by a surface modification step. In some embodiments,the surface modification step is selected from the group including aplasma treatment, a chemical treatment, and an adsorption process. Insome embodiments, the adsorption process includes adsorbing moleculesselected from the group including a polyelectrolyte, apoly(vinylalcohol), an alkylhalosilane, and a ligand.

In some embodiments, the method includes positioning the patternedtemplate and the substrate in a spaced relationship to each other suchthat the patterned template surface and the substrate face each other ina predetermined alignment.

In some embodiments, the disposing of the volume of liquid material onone of the patterned template or the substrate is regulated by aspreading process. In some embodiments, the spreading process includes:

-   -   (a) disposing a first volume of liquid material on one of the        patterned template and the substrate to form a layer of liquid        material thereon; and    -   (b) drawing an implement across the layer of liquid material to:        -   (i) remove a second volume of liquid material from the layer            of liquid material on the one of the patterned template and            the substrate; and        -   (ii) leave a third volume of liquid material on the one of            the patterned template and the substrate.

In some embodiments, an article is contacted with the layer of liquidmaterial and a force is applied to the article to thereby remove theliquid material from the one of the patterned material and thesubstrate. In some embodiments, the article is selected from the groupincluding a roller, a “squeegee” blade type device, a nonplanarpolymeric pad, combinations thereof, or the like. In some embodiments,the liquid material is removed by some other mechanical apparatus.

In some embodiments, the contacting of the patterned template surfacewith the substrate forces essentially all of the disposed liquidmaterial from between the patterned template surface and the substrate.

In some embodiments, the treating of the liquid material includes aprocess selected from the group including a thermal process, a phasechange, an evaporative process, a photochemical process, and a chemicalprocess.

In some embodiments as described in detail herein below, the methodfurther includes:

-   -   (a) reducing the volume of the liquid material disposed in the        plurality of recessed areas by one of:        -   (i) applying a contact pressure to the patterned template            surface; and        -   (ii) allowing a second volume of the liquid to evaporate or            permeate through the template;    -   (b) removing the contact pressure applied to the patterned        template surface;    -   (c) introducing gas within the recessed areas of the patterned        template surface;    -   (d) treating the liquid material to form one or more particles        within the recessed areas of the patterned template surface; and    -   (e) releasing the one or more particles.

In some embodiments, the releasing of the one or more particles isperformed by at least one of:

-   -   (a) applying the patterned template to a substrate, wherein the        substrate has an affinity for the one or more particles;    -   (b) deforming the patterned template such that the one or more        particles is released from the patterned template;    -   (c) swelling the patterned template with a first solvent to        extrude the one or more particles;    -   (d) washing the patterned template with a second solvent,        wherein the second solvent has an affinity for the one or more        particles;    -   (e) applying a mechanical force to the one or more particles;    -   (f) applying the patterned template to a liquid that when        hardened physically entraps particles; and    -   (g) applying the patterned template to a material that when        hardened has a chemical and/or physical interaction with        particles.        In some embodiments, the mechanical force is applied by        contacting one of a Doctor blade and a brush with the one or        more particles. In some embodiments, the mechanical force is        applied by ultrasonics, megasonics, electrostatics, or magnetics        means.

In some embodiments, the method includes harvesting or collecting theparticles. In some embodiments, the harvesting or collecting of theparticles includes a process selected from the group including scrapingwith a doctor blade, a brushing process, a dissolution process, anultrasound process, a megasonics process, an electrostatic process, anda magnetic process. In some embodiments, the harvesting or collecting ofthe particles includes applying a material to at least a portion of asurface of the particle wherein the material has an affinity for theparticles. In some embodiments, the material includes an adhesive orsticky surface. In some embodiments, the material includes, withoutlimitation, one or more of a carbohydrate, an epoxy, a wax, polyvinylalcohol, polyvinyl pyrrolidone, polybutyl acrylate, a polycyanoacrylate, a polyacrylic acid and polymethyl methacrylate. In someembodiments, the harvesting or collecting of the particles includescooling water to form ice (e.g., in contact with the particles). In someembodiments, the presently disclosed subject matter describes a particleor plurality of particles formed by the methods described herein. Insome embodiments, the plurality of particles includes a plurality ofmonodisperse particles. In some embodiments, the particle or pluralityof particles is selected from the group including a semiconductordevice, a crystal, a drug delivery vector, a gene delivery vector, adisease detecting device, a disease locating device, a photovoltaicdevice, a porogen, a cosmetic, an electret, an additive, a catalyst, asensor, a detoxifying agent, an abrasive, such as a CMP, amicro-electro-mechanical system (MEMS), a cellular scaffold, a taggart,a pharmaceutical agent, and a biomarker. In some embodiments, theparticle or plurality of particles include a freestanding structure.

Further, in some embodiments, the presently disclosed subject matterdescribes a method of fabricating isolated liquid objects, the methodincluding (a) contacting a liquid material with the surface of a firstlow surface energy material; (b) contacting the surface of a second lowsurface energy material with the liquid, wherein at least one of thesurfaces of either the first or second low surface energy material ispatterned; (c) sealing the surfaces of the first and the second lowsurface energy materials together; and (d) separating the two lowsurface energy materials to produce a replica pattern including liquiddroplets.

In some embodiments, the liquid material includes poly(ethyleneglycol)-diacrylate. In some embodiments, the low surface energy materialincludes perfluoropolyether-diacrylate. In some embodiments, a chemicalprocess is used to seal the surfaces of the first and the second lowsurface energy materials. In some embodiments, a physical process isused to seal the surfaces of the first and the second low surface energymaterials. In some embodiments, one of the surfaces of the low surfaceenergy material is patterned. In some embodiments, one of the surfacesof the low surface energy material is not patterned.

In some embodiments, the method further includes using the replicapattern composed of liquid droplets to fabricate other objects. In someembodiments, the replica pattern of liquid droplets is formed on thesurface of the low surface energy material that is not patterned. Insome embodiments, the liquid droplets undergo direct or partialsolidification. In some embodiments, the liquid droplets undergo achemical transformation. In some embodiments, the solidification of theliquid droplets or the chemical transformation of the liquid dropletsproduce freestanding objects. In some embodiments, the freestandingobjects are harvested. In some embodiments, the freestanding objects arebonded in place. In some embodiments, the freestanding objects aredirectly solidified, partially solidified, or chemically transformed.

In some embodiments, the liquid droplets are directly solidified,partially solidified, or chemically transformed on or in the patternedtemplate to produce objects embedded in the recesses of the patternedtemplate. In some embodiments, the embedded objects are harvested. Insome embodiments, the embedded objects are bonded in place. In someembodiments, the embedded objects are used in other fabricationprocesses.

In some embodiments, the replica pattern of liquid droplets istransferred to other surfaces. In some embodiments, the transfer takesplace before the solidification or chemical transformation process. Insome embodiments, the transfer takes place after the solidification orchemical transformation process. In some embodiments, the surface towhich the replica pattern of liquid droplets is transferred is selectedfrom the group including a non-low surface energy surface, a low surfaceenergy surface, a functionalized surface, and a sacrificial surface. Insome embodiments, the method produces a pattern on a surface that isessentially free of one or more scum layers. In some embodiments, themethod is used to fabricate semiconductors and other electronic andphotonic devices or arrays. In some embodiments, the method is used tocreate freestanding objects. In some embodiments, the method is used tocreate three-dimensional objects using multiple patterning steps. Insome embodiments, the isolated or patterned object includes materialsselected from the group including organic, inorganic, polymeric, andbiological materials. In some embodiments, a surface adhesive agent isused to anchor the isolated structures on a surface.

In some embodiments, the liquid droplet arrays or solid arrays onpatterned or non-patterned surfaces are used as regiospecific deliverydevices or reaction vessels for additional chemical processing steps. Insome embodiments, the additional chemical processing steps are selectedfrom the group including printing of organic, inorganic, polymeric,biological, and catalytic systems onto surfaces; synthesis of organic,inorganic, polymeric, biological materials; and other applications inwhich localized delivery of materials to surfaces is desired.Applications of the presently disclosed subject matter include, but arenot limited to, micro and nanoscale patterning or printing of materials.In some embodiments, the materials to be patterned or printed areselected from the group including surface-binding molecules, inorganiccompounds, organic compounds, polymers, biological molecules,nanoparticles, viruses, biological arrays, and the like.

In some embodiments, the applications of the presently disclosed subjectmatter include, but are not limited to, the synthesis of polymerbrushes, catalyst patterning for CVD carbon nanotube growth, cellscaffold fabrication, the application of patterned sacrificial layers,such as etch resists, and the combinatorial fabrication of organic,inorganic, polymeric, and biological arrays.

In some embodiments, non-wetting imprint lithography, and relatedtechniques, are combined with methods to control the location andorientation of chemical components within an individual object. In someembodiments, such methods improve the performance of an object byrationally structuring the object so that it is optimized for aparticular application. In some embodiments, the method includesincorporating biological targeting agents into particles for drugdelivery, vaccination, and other applications. In some embodiments, themethod includes designing the particles to include a specific biologicalrecognition motif. In some embodiments, the biological recognition motifincludes biotin/avidin and/or other proteins.

In some embodiments, the method includes tailoring the chemicalcomposition of these materials and controlling the reaction conditions,whereby it is then possible to organize the biorecognition motifs sothat the efficacy of the particle is optimized. In some embodiments, theparticles are designed and synthesized so that recognition elements arelocated on the surface of the particle in such a way to be accessible tocellular binding sites, wherein the core of the particle is preserved tocontain bioactive agents, such as therapeutic molecules. In someembodiments, a non-wetting imprint lithography method is used tofabricate the objects, wherein the objects are optimized for aparticular application by incorporating functional motifs, such asbiorecognition agents, into the object composition. In some embodiments,the method further includes controlling the microscale and nanoscalestructure of the object by using methods selected from the groupincluding self-assembly, stepwise fabrication procedures, reactionconditions, chemical composition, crosslinking, branching, hydrogenbonding, ionic interactions, covalent interactions, and the like. Insome embodiments, the method further includes controlling the microscaleand nanoscale structure of the object by incorporating chemicallyorganized precursors into the object. In some embodiments, thechemically organized precursors are selected from the group includingblock copolymers and core-shell structures.

In sum, the presently disclosed subject matter describes a non-wettingimprint lithography technique that is scalable and offers a simple,direct route to such particles without the use of self-assembled,difficult to fabricate block copolymers and other systems

II.A. Micro and Nano Particles

According to some embodiments of the presently disclosed subject matter,a particle is formed that has a solid geometric shape corresponding to amold having a desired shape and is less than about 10 μm in a givendimension (e.g. minimum, intermediate, or maximum dimension). Theparticle can be of an organic material or an inorganic material and canbe one uniform compound or component or a mixture of compounds orcomponents.

In some embodiments, the particle includes a therapeutic or diagnosticagent coupled with the particle. The therapeutic or diagnostic agent canbe physically coupled or chemically coupled with the particle,encompassed within the particle, at least partially encompassed withinthe particle, coupled to the exterior of the particle, combinationsthereof, and the like. The therapeutic agent can be a drug, a biologic,a ligand, an oligopeptide, a cancer treating agent, a viral treatingagent, a bacterial treating agent, a fungal treating agent, combinationsthereof, or the like.

According to some embodiments, the particle is hydrophilic such that theparticle avoids clearance by biological organism, such as a human.

According to other embodiments, the particle can be substantiallycoated. The coating, for example, can be a sugar based coating where thesugar is preferably glucose, sucrose, maltose, derivatives thereof,combinations thereof, or the like.

In yet other embodiments, the particle can include a functional locationsuch that the particle can be used as an analytical material. Accordingto such embodiments, a particle includes a functional molecular imprint.The functional molecular imprint can include functional monomersarranged as a negative image of a template. The template, for example,can be but is not limited to, an enzyme, a protein, an antibiotic, anantigen, a nucleotide sequence, an amino acid, a drug, a biologic,nucleic acid, combinations thereof, or the like. In other embodiments,the particle itself, for example, can be, but is not limited to, anartificial functional molecule. In one embodiment, the artificialfunctional molecule is a functionalized particle that has been moldedfrom a molecular imprint. As such, a molecular imprint is generated inaccordance with methods and materials of the presently disclosed subjectmatter and then a particle is formed from the molecular imprint, inaccordance with further methods and materials of the presently disclosedsubject matter. Such an artificial functional molecule includessubstantially similar steric and chemical properties of a molecularimprint template. In one embodiment, the functional monomers of thefunctionalized particle are arranged substantially as a negative imageof functional groups of the molecular imprint.

According to some embodiments, particles formed in the patternedtemplates described herein are less than about 10 μm in a dimension. Inother embodiments, the particle is between about 10 μm and about 1μm indimension. In yet further embodiments, the particle is less than about 1μm in dimension. According to some embodiments the particle is betweenabout 1 nm and about 500 nm in a dimension. According to otherembodiments, the particle is between about 10 nm and about 200 nm in adimension. In still further embodiments, the particle is between about80 nm and 120 nm in a dimension. According to still more embodiments theparticle is between about 20 nm and about 120 nm in dimension. Infurther embodiments, the particle has a maximum cross-sectionaldimension of less than about 1 micrometer. In some embodiments, theparticle has a maximum cross-sectional dimension between about 5nanometers and about 1 micrometer. In some embodiments, the particle hasa maximum cross-sectional dimension between about 10 nanometers andabout 1 micrometer. In some embodiments, the particle has a maximumcross-sectional dimension less than about 800 nanometers. In someembodiments, the particle has a maximum cross-sectional dimension lessthan about 750 nanometers. In some embodiments, the particle has amaximum cross-sectional dimension less than about 500 nanometers. Insome embodiments, the particle has a maximum cross-sectional dimensionless than about 300 nanometers. In some embodiments, the particle has amaximum cross-sectional dimension less than about 250 nanometers. Insome embodiments, the particle has a maximum cross-sectional dimensionless than about 200 nanometers. In some embodiments, the particle has amaximum cross-sectional dimension less than about 150 nanometers. Insome embodiments, the particle has a maximum cross-sectional dimensionless than about 100 nanometers. In some embodiments, particles arefabricated in an array of substantially congruent recesses therebyforming a plurality of substantially congruent particles. The dimensionof the particle can be a predetermined dimension, a cross-sectionaldiameter, a circumferential dimension, or the like.

According to further embodiments, the particles include patternedfeatures that are about 2 nm in a dimension. In still furtherembodiments, the patterned features are between about 2 nm and about 200nm. In other embodiments, the particle is less than about 80 nm in awidest dimension.

According to other embodiments, the particles produced by the methodsand materials of the presently disclosed subject matter have a polydispersion index of between about 0.80 and about 1.20, between about0.90 and about 1.10, between about 0.95 and about 1.05, between about0.99 and about 1.01, between about 0.999 and about 1.001, combinationsthereof, and the like. Furthermore, in other embodiments the particlehas a monodispersity.

According to other embodiments, particles of many predetermined regularand irregular shape and size configurations can be made with thematerials and methods of the presently disclosed subject matter.Examples of representative particle shapes that can be made using thematerials and methods of the presently disclosed subject matter include,but are not limited to, non-spherical, spherical, viral shaped, bacteriashaped, cell shaped, rod shaped (e.g., where the rod is less than about200 nm in diameter), chiral shaped, right triangle shaped, flat shaped(e.g., with a thickness of about 2 nm, disc shaped with a thickness ofgreater than about 2 nm, or the like), boomerang shaped, combinationsthereof, and the like.

In some embodiments, the material from which the particles are formedincludes, without limitation, one or more of a polymer, a liquidpolymer, a solution, a monomer, a plurality of monomers, apolymerization initiator, a polymerization catalyst, an inorganicprecursor, an organic material, a natural product, a metal precursor, apharmaceutical agent, a tag, a magnetic material, a paramagneticmaterial, a ligand, a cell penetrating peptide, a porogen, a surfactant,a plurality of immiscible liquids, a solvent, a charged species,combinations thereof, or the like. In further embodiments, the materialsincluded in the particles or from which the particles can be formedincludes compositions, monomers, polymers, and materials found inPolymer Handbook, Grulke, Eric A., et al., John Wiley & Sons; 4thedition (May 29, 2003) which is incorporated herein by reference in itsentirety.

In some embodiments, the monomer includes butadienes, styrenes, propene,acrylates, methacrylates, vinyl ketones, vinyl esters, vinyl acetates,vinyl chlorides, vinyl fluorides, vinyl ethers, acrylonitrile,methacrylnitrile, acrylamide, methacrylamide allyl acetates, fumarates,maleates, ethylenes, propylenes, tetrafluoroethylene, ethers,isobutylene, fumaronitrile, vinyl alcohols, acrylic acids, amides,carbohydrates, esters, urethanes, siloxanes, formaldehyde, phenol, urea,melamine, isoprene, isocyanates, epoxides, bisphenol A, alcohols,chlorosilanes, dihalides, dienes, alkyl olefins, ketones, aldehydes,vinylidene chloride, anhydrides, saccharide, acetylenes, naphthalenes,pyridines, lactams, lactones, acetals, thiiranes, episulfide, peptides,derivatives thereof, and combinations thereof.

In yet other embodiments, the polymer includes polyamides, proteins,polyesters, polystyrene, polyethers, polyketones, polysulfones,polyurethanes, polysiloxanes, polysilanes, cellulose, amylose,polyacetals, polyethylene, glycols, poly(acrylate)s,poly(methacrylate)s, poly(vinyl alcohol), poly(vinylidene chloride),poly(vinyl acetate), poly(ethylene glycol), polystyrene, polyisoprene,polyisobutylenes, poly(vinyl chloride), poly(propylene), poly(lacticacid), polyisocyanates, polycarbonates, alkyds, phenolics, epoxy resins,polysulfides, polyimides, liquid crystal polymers, heterocyclicpolymers, polypeptides, conducting polymers including polyacetylene,polyquinoline, polyaniline, polypyrrole, polythiophene, andpoly(p-phenylene), dendimers, fluoropolymers, derivatives thereof,combinations thereof, and the like.

In still further embodiments, the material from which the particles areformed includes a non-wetting agent. According to another embodiment,the material is a liquid material in a single phase. In otherembodiments, the liquid material includes a plurality of phases. In someembodiments, the liquid material includes, without limitation, one ormore of multiple liquids, multiple immiscible liquids, surfactants,dispersions, emulsions, microemulsions, micelles, particulates,colloids, porogens, active ingredients, combinations thereof, or thelike.

In some embodiments, additional components are included with thematerial of the particle to functionalize the particle. According tothese embodiments the additional components can be encased within theisolated structures, partially encased within the isolated structures,on the exterior surface of the isolated structures, combinationsthereof, or the like. Additional components can include, but are notlimited to, drugs, biologics, more than one drug, more than onebiologic, combinations thereof, and the like.

In some embodiments, the drug is a psychotherapeutic agent. In otherembodiments, the psychotherapeutic agent is used to treat depression andcan include, for example, sertraline, venlafaxine hydrochloride,paroxetine, bupropion, citalopram, fluoxetine, mirtazapine,escitalopram, and the like. In some embodiments, the psychotherapeuticagent is used to treat schizophrenia and can include, for example,olanazapine, risperidone, quetiapine, aripiprazole, ziprasidone, and thelike. According to other embodiments, the psychotherapeutic agent isused to treat attention deficit disorder (ADD) or attention deficithyperactivity disorder (ADHD), and can include, for example,methylphenidate, atomoxetine, amphetamine, dextroamphetamine, and thelike. In some other embodiments, the drug is a cholesterol drug and caninclude, for example, atorvastatin, simvastatin, pravastatin, ezetimibe,rosuvastatin, fenofibrate fluvastatin, and the like. In yet some otherembodiments, the drug is a cardiovascular drug and can include, forexample, amlodipine, valsartan, losartan, hydrochlorothiazide,metoprolol, candesartan, ramipril, irbesartan, amlodipine, benazepril,nifedipine, carvedilol, enalapril, telemisartan, quinapril, doxazosinmesylate, felodipine, lisinopril, and the like. In some embodiments, thedrug is a blood modifier and can include, for example, epoetin alfa,darbepoetin alfa, epoetin beta, clopidogrel, pegfilgrastim, filgrastim,enoxaparin, Factor VIIA, antihemophilic factor, immune globulin, and thelike. According to a further embodiment, the drug can include acombination of the above listed drugs.

In some embodiments, the additional components included with theparticles of the presently disclosed subject matter can include, but arenot limited, to anti-infective agents. In some embodiments, theanti-infective agent is used to treat bacterial infections and caninclude, for example, azithromycin, amoxicillin, clavulanic acid,levofloxacin, clarithromycin, ceftriaxone, ciprofloxacin, piperacillin,tazobactam sodium, imipenem, cilastatin, linezolid, meropenem,cefuroxime, moxifloxacin, and the like. In some embodiments theanti-infective agent is used to treat viral infections and can include,for example, lamivudine, zidovudine, valacyclovir, peginterferon,lopinavir, ritonavir, tenofovir, efavirenz, abacavir, lamivudine,zidovudine, atazanavir, and the like. In other embodiments, theanti-infective agent is used to treat fungal infections and can include,for example, terbinafine, fluconazole, itraconazole, caspofunginacetate, and the like. In some embodiments, the drug is agastrointestinal drug and can include, for example, esomeprazole,lansoprazole, omeprazole, pantoprazole, rabeprazole, ranitidine,ondansetron, and the like. According to yet other embodiments, the drugis a respiratory drug and can include, for example, fluticasone,salmeterol, montelukast, budesonide, formoterol, fexofenadine,cetirizine, desloratadine, mometasone furoate, tiotropium, albuterol,ipratropium, palivizumab, and the like. In yet other embodiments, thedrug is an antiarthritic drug and can include, for example, celecoxib,infliximab, etanercept, rofecoxib, valdecoxib, adalimumab, meloxicam,diclofenac, fentanyl, and the like. According to a further embodiment,the drug can include a combination of the above listed drugs.

According to alternative embodiments, the additional components includedwith the particles of the presently disclosed subject matter caninclude, but are not limited to an anticancer agent and can include, forexample, nitrogen mustard, cisplatin, doxorubicin, docetaxel,anastrozole, trastuzumab, capecitabine, letrozole, leuprolide,bicalutamide, goserelin, rituximab, oxaliplatin, bevacizumab,irinotecan, paclitaxel, carboplatin, imatinib, gemcitabine,temozolomide, gefitinib, and the like. In some embodiments, the drug isa diabetes drug and can include, for example, rosiglitazone,pioglitazone, insulin, glimepiride, voglibose, and the like. In otherembodiments, the drug is an anticonvulsant and can include, for example,gabapentin, topiramate, oxcarbazepine, carbamazepine, lamotrigine,divalproex, levetiracetam, and the like. In some embodiments, the drugis a bone metabolism regulator and can include, for example,alendronate, raloxifene, risedronate, zoledronic, and the like. In someembodiments, the drug is a multiple sclerosis drug and can include, forexample, interferon, glatiramer, copolymer-1, and the like. In otherembodiments, the drug is a hormone and can include, for example,somatropin, norelgestromin, norethindrone, desogestrel, progestin,estrogen, octreotide, levothyroxine, and the like. In yet otherembodiments, the drug is a urinary tract agent, and can include, forexample, tamsulosin, finasteride, tolterodine, and the like. In someembodiments, the drug is an immunosuppressant and can include, forexample, mycophenolate mofetil, cyclosporine, tacrolimus, and the like.In some embodiments, the drug is an ophthalmic product and can include,for example, latanoprost, dorzolamide, botulinum, verteporfin, and thelike. In some embodiments, the drug is a vaccine and can include, forexample, pneumococcal, hepatitis, influenza, diphtheria, and the like.In other embodiments, the drug is a sedative and can include, forexample, zolpidem, zaleplon, eszopiclone, and the like. In someembodiments, the drug is an Alzheimer disease therapy and can include,for example, donepexil, rivastigmine, tacrine, and the like. In someembodiments, the drug is a sexual dysfunction therapy and can include,for example, sildenafil, tadalafil, alprostadil, levothyroxine, and thelike. In an alternative embodiment, the drug is an anesthetic and caninclude, for example, sevoflurane, propofol, mepivacaine, bupivacaine,ropivacaine, lidocaine, nesacaine, etidocaine, and the like. In someembodiments, the drug is a migraine drug and can include, for example,sumatriptan, almotriptan, rizatriptan, naratriptan, and the like. Insome embodiments, the drug is an infertility agent and can include, forexample, follitropin, choriogonadotropin, menotropin, folliclestimulating hormone (FSH), and the like. In some embodiments, the drugis a weight control product and can include, for example, orlistat,dexfenfluramine, sibutramine, and the like. According to a furtherembodiment, the drug can include a combination of the above listeddrugs.

In some embodiments, one or more additional components are included withthe particles. The additional components can include: targeting ligandssuch as cell-targeting peptides, cell-penetrating peptides, integrinreceptor peptide (GRGDSP), melanocyte stimulating hormone, vasoactiveintestional peptide, anti-Her2 mouse antibodies, and the like; vitamins;viruses; polysaccharides; cyclodextrins; liposomes; proteins; opticalnanoparticles such as CdSe for optical applications; boratenanoparticles to aid in boron neutron capture therapy (BNCT) targets;combinations thereof; and the like.

In some embodiments, imaging agents are included with the particles. Insome embodiments, the imaging agent is an x-ray agent and can include,for example, barium sulfate, ioxaglate meglumine, ioxaglate sodium,diatrizoate meglumine, diatrizoate sodium, ioversol, iothalamatemeglumine, iothalamate sodium, iodixanol, iohexol, iopentol, iomeprol,iopamidol, iotroxate meglumine, iopromide, iotrolan, sodiumamidotrizoate, meglumine amidotrizoate, and the like. In someembodiments, the imaging agent is a MRI agent and can include, forexample, gadopentetate dimeglumine, ferucarbotran, gadoxetic aciddisodium, gadobutrol, gadoteddol, gadobenate dimeglumine, ferumoxsil,gadoversetamide, gadolinium complexes, gadodiamide, mangafodipir, andthe like. In some embodiments, the imaging agent is an ultrasound agentand can include, for example, galactose, palmitic acid, SF₆, and thelike. In some embodiments, the imaging agent is a nuclear agent and caninclude, for example, technetium (Tc99m) tetrofosmin, ioflupane,technetium (Tc99m) depreotide, technetium (Tc99m) exametazime,fluorodeoxyglucose (FDG), samarium (Sm153) lexidronam, technetium(Tc99m) mebrofenin, sodium iodide (I125 and I131), technetium (Tc99m)medronate, technetium (Tc99m) tetrofosmin, technetium (Tc99m)fanolesomab, technetium (Tc99m) mertiatide, technetium (Tc99m)oxidronate, technetium (Tc99m) pentetate, technetium (Tc99m) gluceptate,technetium (Tc99m) albumin, technetium (Tc99m) pyrophosphate, thallous(Tl201) chloride, sodium chromate (Cr51), gallium (Ga67) citrate, indium(In111) pentetreotide, iodinated (I125) albumin, chromic phosphate(P32), sodium phosphate (P32), and the like. According to a furtherembodiment, the agent can include a combination of the above listedagents, drugs, biologics, and the like.

According to other embodiments, one or more other drugs can be includedwith the particles of the presently disclosed subject matter and can befound in Physician's Desk Reference, Thomson Healthcare, 59th Bk&Credition (2004), which is incorporated herein by reference in itsentirety.

In some embodiments, the particles are coated with a patient appealingsubstance to facilitate and encourage consumption of the particles asoral drug delivery vehicles. The particles can be coated orsubstantially coated with a substance (e.g., a food substance) that canmask any taste the particle and/or drug combination might have.According to some embodiments, the particle is coated with a sugar basedsubstance to impart to the particle an appealing sweet taste. Accordingto other embodiments, the particles can be coated with materialsdescribed in relation to the fast-dissolve embodiments described hereinabove.

According to some embodiments, radiotracers and/or radiopharmaceuticalsare included with the particles. Examples of radiotracers and/orradiopharmaceuticals that can be combined with the isolated structuresof the presently disclosed subject matter include, but are not limitedto, [¹⁵O]oxygen, [¹⁵O]carbon monoxide, [¹⁵O]carbon dioxide, [¹⁵O]water,[¹³N]ammonia, [¹⁸F]FDG, [¹⁸F]FMISO, [¹⁸F]MPPF, [¹⁸F]A85380, [¹⁸F]FLT,[¹¹C]SCH23390, [¹¹C]flumazenil, [¹¹C]PK11195, [¹¹C]PIB, [¹¹C]AG1478,[¹¹C]choline, [¹¹C]AG957, [¹⁸F]nitroisatin, [¹⁸F]mustard, combinationsthereof, and the like. In some embodiments elemental isotopes areincluded with the particles. In some embodiments, the isotopes include¹¹C, ¹³N, ¹⁵O, ¹⁸F, ³²P, ⁵¹Cr, ⁵⁷Co, ⁶⁷Ga, ⁸¹Kr, ⁸²Rb, ⁸⁹Sr, ⁹⁹Tc,¹¹¹In, ¹²³I, ¹²⁵I, ¹³¹I, ¹³³Xe, ¹⁵³Sm, ²⁰¹Tl, or the like. According toa further embodiment, the isotope can include a combination of the abovelisted isotopes, and the like. Likewise, the particles can include afluorescent label such that the particle can be identified. Examples offluorescent labeled particles are shown in FIGS. 45 and 46. FIG. 45shows a particle that has been fluorescently labeled and is associatedwith a cell membrane and the particle shown in FIG. 46 is within thecell.

According to still further embodiments, contrast agents can be includedwith the material from which the particles are formed. Adding contrastagents enhances diagnostic imaging of physiologic structures forclinical evaluations and other testing. For example, ultrasound imagingtechniques often involve the use of contrast agents, as contrast agentscan serve to improve the quality and usefulness of images which areobtained with ultrasound. The viability of currently availableultrasound contrast agents and methods involving their use is highlydependent on a variety of factors, including the particular region beingimaged. For example, difficulty is encountered in obtaining usefuldiagnostic images of heart tissue and the surrounding vasculature due,at least in part, to the large volume of blood which flows through thechambers of the heart relative to the volume of blood which flows in theblood vessels of the heart tissue itself. The high volume of bloodflowing through the chambers of the heart can result in insufficientcontrast in ultrasound images of the heart region, especially the hearttissue. The high volume of blood flowing through the chambers of theheart also can produce diagnostic artifacts including, for example,shadowing or darkening, in ultrasound images of the heart. Diagnosticartifacts can be highly undesirable since they can hamper or evenprevent visualization of a region of interest. Thus, in certaincircumstances, diagnostic artifacts can render a diagnostic imagesubstantially unusable.

In addition to ultrasound, computed tomography (CT) is a valuablediagnostic imaging technique for studying various areas of the body.Like ultrasound, CT imaging is greatly enhanced with the aid of contrastagents. In CT, the radiodensity (electron density) of matter ismeasured. Because of the similarity in the measured densities of varioustissues in the body, it has been necessary to use contrast agents whichcan change the relative densities of different tissues. Thischaracteristic has resulted in an overall improvement in the diagnosticefficacy of CT. Barium and iodine compounds, for example, have beendeveloped for this purpose and can be included with the particles of thepresently disclosed subject matter in some embodiments. Accordingly, inother embodiments, contrast agents that can be used with the materialsof the presently disclosed subject matter, include for example, but arenot limited to, barium sulfate, lodinated water-soluble contrast media,combinations thereof, and the like.

Magnetic resonance imaging (MRI) is another diagnostic imaging techniquethat is used for producing cross-sectional images of a tissue in avariety of scanning planes. Like ultrasound and CT, MRI also benefitsfrom the use of contrast agents. In some embodiments of the presentlydisclosed subject matter, contrast agents for MRI are used with thematerials of the presently disclosed subject matter to enhance MRIimaging. Contrast agents for MRI imaging that can be useful with thematerials of the presently disclosed subject matter include, but are notlimited to, paramagnetic contrast agents, metal ions, transition metalions, metal ions that are chelated with ligands, metal oxides, ironoxides, nitroxides, stable free radicals, stable nitroxides, lanthanideand actinide elements, lipophilic derivatives, proteinaceousmacromolecules, alkylated, nitroxides2,2,5,5-tetramethyl-1-pyrrolidinyloxy, free radical,2,2,6,6-tetramethyl-1-piperidinyloxy, free radical, combinationsthereof, and the like.

According to yet other embodiments contrast agents that can be used withthe materials of the presently disclosed subject matter include, but arenot limited to, superparamagnetic contrast agents, ferro- orferrimagnetic compounds such as pure iron, magnetic iron oxide, such asmagnetite, γ-Fe₂O₃, Fe₃O₄, manganese ferrite, cobalt ferrite, nickelferrite; paramagnetic gases such as oxygen 17 gas, hyperpolarized xenon,neon, helium gas, combinations thereof, and the like. If desired, theparamagnetic or superparamagnetic contrast agents used with thematerials of the presently disclosed include, but are not limited to,paramagnetic or superparamagetic agents that are delivered as alkylatedor having other derivatives incorporated into the compositions,combinations thereof, and the like.

In yet another embodiment, contrast agents for X-ray techniques usefulfor combination with the particles of the presently disclosed subjectmatter include, but are not limited to, carboxylic acid and non-ionicamide contrast agents typically containing at least one2,4,6-triiodophenyl group having substituents such as carboxyl,carbamoyl, N-alkylcarbamoyl, N-hydroxyalkylcarbamoyl, acylamino,N-alkylacylamino or acylaminomethyl at the 3- and/or 5-positions, as inmetrizoic acid, diatrizoic acid, iothalamic acid, ioxaglic acid,iohexol, iopentol, iopamidol, iodixanol, iopromide, metrizamide,iodipamide, meglumine iodipamide, meglumine acetrizoate, megluminediatrizoate, combinations thereof, and the like.

Still other contrast agents that can be included with the particlematerials of the presently disclosed subject matter include, but are notlimited to, barium sulfate, a barium sulfate suspension, sodiumbicarbonate and tartaric acid mixtures, lothalamate meglumine,lothalamate sodium, hydroxypropyl methylcellulose, ferumoxsil, ioxaglatemeglumine, ioxaglate sodium, diatrizoate meglumine, diatrizoate sodium,gadoversetamide, ioversol, organically bound iodine, methiodal sodium,ioxitalamate meglumine, iocarmate meglumine, metrizamide, iohexal,iopamidol, combinations thereof, and the like.

U.S. Pat. Nos. 6,884,407 and 6,331,289, along with the references citedtherein, disclose contrasts that are useful with the particles of thepresently disclosed subject matter, these references are incorporated byreference herein along with the references cited therein.

According to further embodiments the particle can include or can beformed into and used as a tag or a taggant. A taggant that can beincluded in the particle or can be the particle includes, but is notlimited to, a fluorescent, radiolabeled, magnetic, biologic, shapespecific, size specific, combinations thereof, or the like.

In some embodiments, a therapeutic agent for combination with theparticles of the presently disclosed subject matter is selected from oneof a drug and genetic material. In some embodiments, the geneticmaterial includes, without limitation, one or more of a non-viral genevector, DNA, RNA, RNAi, a viral particle, agents described elsewhereherein, combinations thereof, or the like.

In some embodiments, the particle includes a biodegradable polymer. Inother embodiments, the polymer is modified to be a biodegradable polymer(e.g., a poly(ethylene glycol) that is functionalized with a disulfidegroup). In some embodiments, the biodegradable polymer includes, withoutlimitation, one or more of a polyester, a polyanhydride, a polyamide, aphosphorous-based polymer, a poly(cyanoacrylate), a polyurethane, apolyorthoester, a polydihydropyran, a polyacetal, combinations thereof,or the like.

In some embodiments, the polyester includes, without limitation, one ormore of polylactic acid, polyglycolic acid, poly(hydroxybutyrate),poly(ε-caprolactone), poly(β-malic acid), poly(dioxanones), combinationsthereof, or the like. In some embodiments, the polyanhydride includes,without limitation, one or more of poly(sebacic acid), poly(adipicacid), poly(terpthalic acid), combinations thereof, or the like. In yetother embodiments, the polyamide includes, without limitation, one ormore of poly(imino carbonates), polyaminoacids, combinations thereof, orthe like.

According to some embodiments, the phosphorous-based polymer includes,without limitation, one or more of a polyphosphate, a polyphosphonate, apolyphosphazene, combinations thereof, or the like. Further, in someembodiments, the biodegradable polymer further includes a polymer thatis responsive to a stimulus. In some embodiments, the stimulus includes,without limitation, one or more of pH, radiation, ionic strength,oxidation, reduction, temperature, an alternating magnetic field, analternating electric field, combinations thereof, or the like. In someembodiments, the stimulus includes an alternating magnetic field.

In some embodiments, a pharmaceutical agent can be combined with theparticle material. The pharmaceutical agent can be, but is not limitedto, a drug, a peptide, RNAi, DNA, combinations thereof, or the like. Inother embodiments, the tag is selected from the group including afluorescence tag, a radiolabeled tag, a contrast agent, combinationsthereof, or the like. In some embodiments, the ligand includes a celltargeting peptide, or the like.

In use, the particles of the presently disclosed subject matter can beused as treatment devices. In such uses, the particle is administered ina therapeutically effective amount to a patient. According to yet otheruses, the particle can be utilized as a physical tag. In such uses, aparticle of a predetermined shape having a diameter of less than about 1μm in a dimension is used as a taggant to identify products or theorigin of a product. The particle as a taggant can be eitheridentifiable to a particular shape or a particular chemical composition.

Further uses of the micro and/or nano particles include medicaltreatments such as orthopedic, oral, maxillofacial, and the like. Forexample, the particles described above that are or includepharmaceutical agents can be used in combination with traditionalhygiene and/or surgical procedures. According to such an application,the particles can be used to directly and locally deliver pharmaceuticalagents, or the like to an area of surgical interest. In someembodiments, medications used in oral medicine can fight oral diseases,prevent or treat infections, control pain, relieve anxiety, assist inthe regeneration of damaged tissue, combinations thereof, and the like.For example, during oral or maxillofacial treatments, bleeding oftenoccurs. As a result, bacteria from the mouth can directly enter thebloodstream and easily reach the heart. This occurrence presents a riskfor some persons with cardiac abnormalities because the bacteria cancause bacterial endocarditis, a serious inflammation of the heart valvesor tissues. Antibiotics reduce this risk. Traditional antibioticdelivery techniques, however, can be slow to reach the bloodstream, thusgiving the bacterial a head start. To the contrary, applying particlesof the presently disclosed subject matter, made from or includingappropriate antibiotics, directly to the site of oral or maxillofacialtreatment can greatly reduce the probability of a serious bacterialinfection. Such procedures aided by the particles can includeprofessional teeth cleaning, incision and drainage of infected oraltissue, oral injections, extractions, surgeries that involve themaxillary sinus, combinations thereof, and the like.

According to further embodiments, compositions can be formulated andmade into particles according to materials and methods of the presentlydisclosed subject matter that are designed to be applied to defectiveteeth and gums for preventing diseases, such as carious tooth, pyorrheaalveolaris, or the like.

Further embodiments include particles having a composition for therepair and healing of tissue, bone defects and bone voids, resins forartificial teeth, resins for tooth bed, and other tooth fillers. Forexample, particles can be constructed from calcium based component, suchas, but not limited to, calcium phosphates, calcium sulfates, calciumcarbonates, calcium bone cements, amorphous calcium phosphate,crystalline calcium phosphate, combinations thereof, and the like. Inuse, such particles can be locally applied to a site of orthopedictreatment to facilitate recovery of the natural bone material.Furthermore, because of the small size of the particles and the abilityto form the particles in practically any shape and configurationdesirable, the particles can be administered to a site of orthopedicinterest and interact with the site on a scale of the particle size.That is, the particles can integrate into very small spaces, cracks,gaps, and the like within the bone, such as a bone fracture, or betweenthe bone and an implant. Thus, the particles can deliver pharmaceutical,regenerative, or the like materials to the orthopedic treatment site andintegrate these materials where they were not previously applyable.Still further, the particles can increase the mechanical strength andintegrity of fixation of a bone implant, such as an artificial jointfixation, because, due to control over the size and shape of theparticles, they can neatly and orderly fill small voids between theimplant and the natural bone tissue.

In other embodiments, medications to control pain and anxiety that arecommonly used in oral, maxillofacial, orthopedic, and other procedurescan be included in the particles. Such agents that can be incorporatedwith the particle include, but are not limited to, anti-inflammatorymedications that are used to relieve the discomfort of mouth and gumproblems, and can include corticosteroids, opioids, carprofen,meloxicam, etodolac, diclofenac, flurbiprofen, ibuprofen, ketorolac,nabumetone, naproxen, naproxen sodium, and oxaprozin. Oral anestheticsare used to relieve pain or irritation caused by many conditions,including toothaches, teething, sores, or dental appliances, and caninclude articaine, epinephrine, ravocaine, novocain, levophed,propoxycaine, procaine, norepinephrine bitartrate, marcaine, lidocaine,carbocaine, neocobefrin, mepivacaine, levonordefrin, etidocaine,dyclonine, and the like. Antibiotics are commonly used to control plaqueand gingivitis in the mouth, treat periodontal disease, as well asreduce the risk of bacteria from the mouth entering the bloodstream.Oral antibiotics can include chlorhexidine, doxycycline, demeclocycline,minocycline, oxytetracycline, tetracycline, triclosan, clindamycin,orfloxacin, metronidazole, tinidazole, and ketoconazole. Fluoride alsocan be or be included in the particles of the presently disclosedsubject matter and is used to prevent tooth decay. Fluoride is absorbedby teeth and helps strengthen teeth to resist acid and block thecavity-forming action of bacteria. As a varnish or a mouth rinse,fluoride helps reduce tooth sensitivity. Other useful agents for dentalapplications are substances such as flavonoids, benzenecarboxylic acids,benzopyrones, steroids, pilocarpine, terpenes, and the like. Stillfurther agents used within the particles include anethole, anisaldehyde,anisic acid, cinnamic acid, asarone, furfuryl alcohol, furfural, cholicacid, oleanolic acid, ursolic acid, sitosterol, cineol, curcumine,alanine, arginine, homocerine, mannitol, berterine, bergapten, santonin,caryophyllene, caryophyllene oxide, terpinene, chymol, terpinol,carvacrol, carvone, sabinene, inulin, lawsone, hesperedin, naringenin,flavone, flavonol, quercetin, apigenin, formonoretin, coumarin, acetylcoumarin, magnolol, honokiol, cappilarin, aloetin, and the like. Stillfurther oral and maxillofacial treatment compounds include sustainedrelease biodegradable compounds, such as, for example (meth)acrylatetype monomers and/or polymers. Other compounds useful for the particlesof the presently disclosed subject matter can be found in U.S. Pat. No.5,006,340, which is incorporated herein by reference in its entirety.

III. Formation of Rounded Particles Through “Liquid Reduction”

Referring now to FIGS. 3A through 3F, the presently disclosed subjectmatter provides a “liquid reduction” process for forming particles thathave shapes that do not conform to the shape of the template, includingbut not limited to spherical and non-spherical, regular and non-regularmicro- and nanoparticles. For example, a “cube-shaped” template canallow for sphereical particles to be made, whereas a “Blockarrow-shaped” template can allow for “lolli-pop” shaped particles orobjects to be made wherein the introduction of a gas allows surfacetension forces to reshape the resident liquid prior to treating it.While not wishing to be bound by any particular theory, the non-wettingcharacteristics that can be provided in some embodiments of thepresently disclosed patterned template and/or treated or coatedsubstrate allows for the generation of rounded, e.g., spherical,particles.

Referring now to FIG. 3A, droplet 302 of a liquid material is disposedon substrate 300, which in some embodiments is coated or treated with anon-wetting material 304. A patterned template 108, which includes aplurality of recessed areas 110 and patterned surface areas 112, also isprovided.

Referring now to FIG. 3B, patterned template 108 is contacted withdroplet 302. The liquid material including droplet 302 then entersrecessed areas 110 of patterned template 108. In some embodiments, aresidual, or “scum,” layer RL of the liquid material including droplet302 remains between the patterned template 108 and substrate 300.

Referring now to FIG. 3C, a first force F_(a1) is applied to patternedtemplate 108. A contact point CP is formed between the patternedtemplate 108 and the substrate and displacing residual layer RL.Particles 306 are formed in the recessed areas 110 of patterned template108.

Referring now to FIG. 3D, a second force F_(a2), wherein the forceapplied by F_(a2) is greater than the force applied by F_(a1), is thenapplied to patterned template 108, thereby forming smaller liquidparticles 308 inside recessed areas 112 and forcing a portion of theliquid material including droplet 302 out of recessed areas 112.

Referring now to FIG. 3E, the second force F_(a2) is released, therebyreturning the contact pressure to the original contact pressure appliedby first force F_(a1). In some embodiments, patterned template 108includes a gas permeable material, which allows a portion of space withrecessed areas 112 to be filled with a gas, such as nitrogen, therebyforming a plurality of liquid spherical droplets 310. Once this liquidreduction is achieved, the plurality of liquid spherical droplets 310are treated by a treating process T_(r).

Referring now to FIG. 3F, treated liquid spherical droplets 310 arereleased from patterned template 108 to provide a plurality offreestanding spherical particles 312.

IIIA. Formation of Small Particles through Evaporation

Referring now to FIGS. 41A through 41E, an embodiment of the presentlydisclosed subject matter includes a process for forming particlesthrough evaporation. In one embodiment, the process produces a particlehaving a shape that does not necessarily conform to the shape of thetemplate. The shape can include, but is not limited to, any threedimensional shape. According to some embodiments, the particle forms aspherical or non-spherical and regular or non-regular shaped micro- andnanoparticle. While not wishing to be bound by any particular theory, anexample of producing a spherical or substantially spherical particleincludes using a patterned template and/or substrate of a non-wettingmaterial or treating the surfaces of the patterned template andsubstrate particle forming recesses with a non-wetting agent such thatthe material from which the particle will be formed does not wet thesurfaces of the recess. Because the material from which the particlewill be formed cannot wet the surfaces of the patterned template and/orsubstrate the particle material has a greater affinity for itself thanthe surfaces of the recesses and thereby forms a rounded, curved, orsubstantially spherical shape.

A non-wetting substance can be defined through the concept of thecontact angle (Θ), which can be used quantitatively to measureinteraction between any liquid and solid surface. When the contact anglebetween a drop of liquid on the surface is 90<Θ3<180, the surface isconsidered non-wetting. In general, fluorinated surfaces are non-wettingto aqueous and organic liquids. Fluorinated surfaces can include afluoropolyether material, a fluoroolefin material, an acrylate material,a silicone material, a styrenic material, a fluorinated thermoplasticelastomer (TPE), a triazine fluoropolymer, a perfluorocyclobutylmaterial, a fluorinated epoxy resin, and/or a fluorinated monomer orfluorinated oligomer that can be polymerized or crosslinked by ametathesis polymerization reaction, surfaces created by treating asilicon or glass surface with a fluorinated silane, or coating a surfacewith a fluorinated polymer. Further, surfaces of materials that aretypically wettable materials can be made non-wettable by surfacetreatments. Materials that can be made substantially non-wetting bysurface treatments include, but are not limited to, a typical wettablepolymer material, an inorganic material, a silicon material, a quartzmaterial, a glass material, combinations thereof, and the like. Surfacetreatments to make these types of materials non-wetting include, forexample, layering the wettable material with a surface layer of theabove described non-wetting materials, and techniques of the like thatwill be appreciated by one of ordinary skill in the art.

Referring now to FIG. 41A, droplet 4102 of a liquid material of thepresently disclosed subject matter that is to become the particle isdisposed on non-wetting substrate 4100, which in some embodiments is amaterial or a surface coated or treated with a non-wetting material, asdescribed herein above. A patterned template 4108, which includes aplurality of recessed areas 4110 and patterned surface areas 4112, alsois provided.

Referring now to FIG. 41B, patterned template 4108 is contacted withdroplet 4102. The liquid material including droplet 4102 then entersrecessed areas 4110 of patterned template 4108. According to someembodiments, mechanical or physical manipulation of droplet 4102 andpatterned template 4108 is provided to facilitate the droplet 4102 insubstantially filling and conforming to recessed areas 4110. Suchmechanical and/or physical manipulation can include, but is not limitedto, vibration, rotation, centrifugation, pressure differences, a vacuumenvironment, combinations thereof, or the like. A contact point CP isformed between the patterned surface areas 4112 and the substrate 4100.Particles 4106 are formed in the recessed areas 4110 of patternedtemplate 4108.

Referring now to FIG. 41C, an evaporative process, E, is performed,thereby reducing the volume of liquid particles 4106 inside recessedareas 4110. Examples of an evaporative process E that can be used withthe present embodiments include forming patterned template 4108 from agas permeable material, which allows volatile components of the materialto become the particles to pass through the template, thereby reducingthe volume of the material to become the particles in the recesses.According to another embodiment, an evaporative process E suitable foruse with the presently disclosed subject matter includes providing aportion of the recessed areas 4110 filled with a gas, such as nitrogen,which thereby increases the evaporation rate of the material to becomethe particles. According to further embodiments, after the recesses arefilled with material to become the particles, a space can be leftbetween the patterned template and substrate such that evaporation isenhanced. In yet another embodiment, the combination of the patternedtemplate, substrate, and material to become the particle can be heatedor otherwise treated to enhance evaporation of the material to becomethe particle. Combinations of the above described evaporation processesare encompassed by the presently disclosed subject matter.

Referring now to FIG. 41D, once liquid reduction is achieved, theplurality of liquid droplets 4114 are treated by a treating processT_(r). Treating process T_(r) can be photo curing, thermal curing, phasechange, solvent evaporation, crystallization, oxidative/reductiveprocesses, combinations thereof, or the like to solidify the material ofdroplet 4102.

Referring now to FIG. 41E, patterned template 4108 is separated fromsubstrate 4100 according to methods and techniques described herein.After separation of patterned template 4108 from substrate 4100, treatedliquid spherical droplets 4114 are released from patterned template 4108to provide a plurality of freestanding spherical particles 4116. In someembodiments release of the particles 4116 is facilitated by a solvent,applying a substance to the particles with an affinity for theparticles, subjecting the particles to gravitational forces,combinations thereof, and the like.

According to some embodiments the particles are less than about 200 nmin diameter. According to some embodiments the particles are betweenabout 80 nm and 200 nm in diameter. According to some embodiments theparticles are between about 100 nm and about 200 nm in diameter.

IV. Formation of Polymeric Nano- to Micro-Electrets

Referring now to FIGS. 4A and 4B, in some embodiments, the presentlydisclosed subject matter describes a method for preparing polymericnano- to micro-electrets by applying an electric field during thepolymerization and/or crystallization step during molding (FIG. 4A) toyield a charged polymeric particle (FIG. 4B). In some embodiments, thecharged polymeric particles spontaneously aggregate into chain-likestructures (FIG. 4D) instead of the random configurations shown in FIG.4C.

In some embodiments, the charged polymeric particle includes a polymericelectret. In some embodiments, the polymeric electret includes apolymeric nano-electret. In some embodiments, the charged polymericparticles aggregate into chain-like structures. In some embodiments, thecharged polymeric particles include an additive for anelectro-rheological device. In some embodiments, the electro-rheologicaldevice is selected from the group including clutches and activedampening devices. In some embodiments, the charged polymeric particlesinclude nano-piezoelectric devices. In some embodiments, thenano-piezoelectric devices are selected from the group includingactuators, switches, and mechanical sensors.

V. Formation of Multilayer Structures

In some embodiments, the presently disclosed subject matter provides amethod for forming multilayer structures, including multilayerparticles. In some embodiments, the multilayer structures, includingmultilayer particles, include nanoscale multilayer structures. In someembodiments, multilayer structures are formed by depositing multiplethin layers of immisible liquids and/or solutions onto a substrate andforming particles as described by any of the methods hereinabove. Theimmiscibility of the liquid can be based on any physical characteristic,including but not limited to density, polarity, and volatility. Examplesof possible morphologies of the presently disclosed subject matter areillustrated in FIGS. 5A-5C and include, but are not limited to,multi-phase sandwich stuctures, core-shell particles, and internalemulsions, microemulsions and/or nano-sized emulsions.

Referring now to FIG. 5A, a multi-phase sandwich structure 500 of thepresently disclosed subject matter is shown, which by way of example,includes a first liquid material 502 and a second liquid material 504.

Referring now to FIG. 5B, a core-shell particle 506 of the presentlydisclosed subject matter is shown, which by way of example, includes afirst liquid material 502 and a second liquid material 504.

Referring now to FIG. 5C, an internal emulsion particle 508 of thepresently disclosed subject matter is shown, which by way of example,includes a first liquid material 502 and a second liquid material 504.

More particularly, in some embodiments, the method includes disposing aplurality of immiscible liquids between the patterned template andsubstrate to form a multilayer structure, e.g., a multilayernanostructure. In some embodiments, the multilayer structure includes amultilayer particle. In some embodiments, the multilayer structureincludes a structure selected from the group including multi-phasesandwich structures, core-shell particles, internal emulsions,microemulsions, and nanosized emulsions.

VI. Fabrication of Complex Multi-Dimensional Structures

In some embodiments, the currently disclosed subject matter provides aprocess for fabricating complex, multi-dimensional structures. In someembodiments, complex multi-dimensional structures can be formed byperforming the steps illustrated in FIGS. 2A-2E. In some embodiments,the method includes imprinting onto a patterned template that is alignedwith a second patterned template (instead of imprinting onto a smoothsubstrate) to generate isolated multidimensional structures that arecured and released as described herein. A schematic illustration of anembodiment of a process for forming complex multi-dimensional structuresand examples of such structures are provided in FIGS. 6A-6C.

Referring now to FIG. 6A, a first patterned template 600 is provided.First patterned template 600 includes a plurality of recessed areas 602and a plurality of non-recessed surfaces 604. Also provided is a secondpatterned template 606. Second patterned template 606 includes aplurality of recessed areas 608 and a plurality of non-recessed surfaces610. As shown in FIG. 6A, first patterned template 600 and secondpatterned template 606 are aligned in a predetermined spacedrelationship. A droplet of liquid material 612 is disposed between firstpatterned template 600 and second patterned template 606.

Referring now to FIG. 6B, patterned template 600 is contacted withpatterned template 606. A force F_(a) is applied to patterned template600 causing the liquid material including droplet 612 to migrate to theplurality of recessed areas 602 and 608. The liquid material includingdroplet 612 is then treated by treating process T_(r) to form apatterned, treated liquid material 614.

Referring now to FIG. 6C, the patterned, treated liquid material 614 ofFIG. 6B is released by any of the releasing methods described herein toprovide a plurality of multi-dimensional patterned structures 616.

In some embodiments, patterned structure 616 includes ananoscale-patterned structure. In some embodiments, patterned structure616 includes a multi-dimensional structure. In some embodiments, themulti-dimensional structure includes a nanoscale multi-dimensionalstructure. In some embodiments, the multi-dimensional structure includesa plurality of structural features. In some embodiments, the structuralfeatures include a plurality of heights.

In some embodiments, a microelectronic device including patternedstructure 616 is provided. Indeed, patterned structure 616 can be anystructure imaginable, including “dual damscene” structures formicroelectronics. In some embodiments, the microelectronic device isselected from the group including integrated circuits, semiconductorparticles, quantum dots, and dual damascene structures. In someembodiments, the microelectronic device exhibits certain physicalproperties selected from the group including etch resistance, lowdielectric constant, high dielectric constant, conducting,semiconducting, insulating, porosity, and non-porosity.

In some embodiments, the presently disclosed subject matter discloses amethod of preparing a multidimensional, complex structure. Referring nowto FIGS. 7A-7F, in some embodiments, a first patterned template 700 isprovided. First patterned template 700 includes a plurality ofnon-recessed surface areas 702 and a plurality of recessed surface areas704. Continuing particularly with FIG. 7A, also provided is a substrate706. In some embodiments, substrate 706 is coated with a non-wettingagent 708. A droplet of a first liquid material 710 is disposed onsubstrate 706.

Referring now to FIGS. 7B and 7C, first patterned template 700 iscontacted with substrate 706. A force F_(a) is applied to firstpatterned template 700 such that the droplet of the first liquidmaterial 710 is forced into recesses 704. The liquid material includingthe droplet of first liquid material 710 is treated by a first treatingprocess T_(r1) to form a treated first liquid material within theplurality of recesses 704. In some embodiments, first treating processT_(r1) includes a partial curing process causing the treated firstliquid material to adhere to substrate 706. Referring particularly toFIG. 7C, first patterned template 700 is removed to provide a pluralityof structural features 712 on substrate 706.

Referring now to FIGS. 7D-7F, a second patterned template 714 isprovided. Second patterned substrate 714 includes a plurality ofrecesses 716, which are filled with a second liquid material 718. Thefilling of recesses 716 can be accomplished in a manner similar to thatdescribed in FIGS. 7A and 7B with respect to recesses 704. Referringparticularly to FIG. 7E, second patterned template 714 is contacted withstructural features 712. Second liquid material 718 is treated with asecond treating process T_(r2) such that the second liquid material 718adheres to the plurality of structural feature 712, thereby forming amultidimensional structure 720. Referring particularly to FIG. 7F,second patterned template 714 and substrate 706 are removed, providing aplurality of free standing multidimensional structures 722. In someembodiments, the process schematically presented in FIGS. 7A-7F can becarried out multiple times as desired to form intricate nanostructures.

Accordingly, in some embodiments, a method for forming multidimensionalstructures is provided, the method including:

-   -   (a) providing a particle prepared by the process described in        the figures;    -   (b) providing a second patterned template;    -   (c) disposing a second liquid material in the second patterned        template;    -   (d) contacting the second patterned template with the particle        of step (a); and    -   (e) treating the second liquid material to form a        multidimensional structure.        VII. Functionalization of Particles

In some embodiments, the presently disclosed subject matter provides amethod for functionalizing isolated micro- and/or nanoparticles. In oneembodiment, the functionalization includes introducing chemicalfunctional groups to a surface either physically or chemically. In someembodiments, the method of functionalization includes introducing atleast one chemical functional group to at least a portion ofmicroparticles and/or nanoparticles. In some embodiments, particles 3605are at least partially functionalized while particles 3605 are incontact with an article 3600. In one embodiment, the particles 3605 tobe functionalized are located within a mold or patterned template 108(FIGS. 35A-36D). In some embodiments, particles 3605 to befunctionalized are attached to a substrate (e.g., substrate 4010 ofFIGS. 40A-40D). In some embodiments, at least a portion of the exteriorof the particles 3605 can be chemically modified by performing the stepsillustrated in FIGS. 36A-36D. In one embodiment, the particles 3605 tobe functionalized are located within article 3600 as illustrated inFIGS. 36A and 40A. As illustrated in FIGS. 36A-36D and 40A-40D, someembodiments include contacting an article 3600 containing particles 3605with a solution 3602 containing a modifying agent 3604.

In one embodiment, illustrated in FIGS. 36C and 40C, modifying agent3604 attaches (e.g., chemically) to exposed particle surface 3606 bychemically reacting with or physically adsorbing to a linker group onparticle surface 3606. In one embodiment, the linker group on particle3606 is a chemical functional group that can attach to other species viachemical bond formation or physical affinity. In some embodiments, thelinker group includes a functional group that includes, withoutlimitation, sulfides, amines, carboxylic acids, acid chlorides,alcohols, alkenes, alkyl halides, isocyanates, compounds disclosedelsewhere herein, combinations thereof, or the like.

In one embodiment, illustrated in FIGS. 36D and 40D, excess solution isremoved from article 3600 while particle 3605 remains in contact witharticle 3600. In some embodiments, excess solution is removed from thesurface containing the particles. In some embodiments, excess solutionis removed by rinsing with or soaking in a liquid, by applying an airstream, or by physically shaking or scraping the surface. In someembodiments, the modifying agent includes an agent selected from thegroup including dyes, fluorescent tags, radiolabeled tags, contrastagents, ligands, peptides, pharmaceutical agents, proteins, DNA, RNA,siRNA, compounds and materials disclosed elsewhere herein, combinationsthereof, and the like.

In one embodiment, functionalized particles 3608, 4008 are harvestedfrom article 3600 using, for example, methods described herein. In someembodiments, functionalizing and subsequently harvesting particles thatreside on an article (e.g., a substrate, a mold or patterned template)have advantages over other methods (e.g., methods in which the particlesmust be functionalized while in solution). In one embodiment of thepresently disclosed subject matter, fewer particles are lost in theprocess, giving a high product yield. In one embodiment of the presentlydisclosed subject matter, a more concentrated solution of the modifyingagent can be applied in lower volumes. In one embodiment of thepresently disclosed subject matter, where particles are functionalizedwhile they remain associated with article 3600 functionalization doesnot need to occur in a dilute solution. In one embodiment, the use ofmore concentrated solution facilitates, for example, the use of lowervolumes of modifying agent and/or lower times to functionalize. In oneembodiment, particles in a tight, 2-dimensional array, but not touching,are susceptible to application of thin, concentrated solutions forfaster functionalization. In some embodiments, lower volume/higherconcentration modifying agent solutions are useful, for example, inconnection with modifying agents that are difficult and expensive tomake and handle (e.g., biological agents such as peptides, DNA, or RNA).In some embodiments, functionalizing particles that remain connected toarticle 3600 eliminates difficult and/or time-consuming steps to removeexcess unreacted material (e.g., dialysis, extraction, filtration andcolumn separation). In one embodiment of the presently disclosed subjectmatter, highly pure functionalized product can be produced at a reducedeffort and cost.

VII. Imprint Lithography

Referring now to FIGS. 8A-8D, a method for forming a pattern on asubstrate is illustrated. In the embodiment illustrated in FIG. 8, animprint lithography technique is used to form a pattern on a substrate.

Referring now to FIG. 8A, a patterned template 810 is provided. In someembodiments, patterned template 810 includes a solvent resistant, lowsurface energy polymeric material, derived from casting low viscosityliquid materials onto a master template and then curing the lowviscosity liquid materials to generate a patterned template as definedhereinabove. Patterned template 810 further includes a first patternedtemplate surface 812 and a second template surface 814: The firstpatterned template surface 812 further includes a plurality of recesses816. The patterned template derived from a solvent resistant, lowsurface energy polymeric material could be mounted on another materialto facilitate alignment of the patterned template or to facilitatecontinuous processing such as a conveyor belt. This might beparticularly useful in the fabrication of precisely placed structures ona surface, such as in the fabrication of a complex devices or asemiconductor, electronic or photonic devices.

Referring again to FIG. 8A, a substrate 820 is provided. Substrate 820includes a substrate surface 822. In some embodiments, substrate 820 isselected from the group including a polymer material, an inorganicmaterial, a silicon material, a quartz material, a glass material, andsurface treated variants thereof. In some embodiments, at least one ofpatterned template 810 and substrate 820 has a surface energy lower than18 mN/m. In some embodiments, at least one of patterned template 810 andsubstrate 820 has a surface energy lower than 15 mN/m. According to afurther embodiment the patterned template 810 and/or the substrate 820has a surface energy between about 10 mN/m and about 20 mN/m. Accordingto some embodiments, the patterned template 810 and/or the substrate 820has a low surface energy of between about 12 mN/m and about 15 mN/m.

In some embodiments, as illustrated in FIG. 8A, patterned template 810and substrate 820 are positioned in a spaced relationship to each othersuch that first patterned template surface 812 faces substrate surface822 and a gap 830 is created between first patterned template surface812 and substrate surface 822. This is an example of a predeterminedrelationship.

Referring now to FIG. 8B, a volume of liquid material 840 is disposed ingap 830 between first patterned template surface 812 and substratesurface 822. In some embodiments, the volume of liquid material 840 isdisposed directed on a non-wetting agent (not shown), which is disposedon first patterned template surface 812.

Referring now to FIG. 8C, in some embodiments, first patterned template812 is contacted with the volume of liquid material 840. A force F_(a)is applied to second template surface 814 thereby forcing the volume ofliquid material 840 into the plurality of recesses 816. In someembodiments, as illustrated in FIG. 8C, a portion of the volume ofliquid material 840 remains between first patterned template surface 812and substrate surface 820 after force F_(a) is applied.

Referring again to FIG. 8C, in some embodiments, the volume of liquidmaterial 840 is treated by a treating process T_(r) while force F_(a) isbeing applied to form a treated liquid material 842. In someembodiments, treating process T_(r) includes a process selected from thegroup including a thermal process, a photochemical process, and achemical process.

Referring now to FIG. 8D, a force F_(r) is applied to patterned template810 to remove patterned template 810 from treated liquid material 842 toreveal a pattern 850 on substrate 820 as shown in FIG. 8E. In someembodiments, a residual, or “scum,” layer 852 of treated liquid material842 remains on substrate 820.

More particularly, the method for forming a pattern on a substrateincludes:

-   -   (a) providing patterned template and a substrate, wherein the        patterned template includes a patterned template surface having        a plurality of recessed areas formed therein;    -   (b) disposing a volume of liquid material in or on at least one        of:        -   (i) the patterned template surface;        -   (ii) the plurality of recessed areas; and        -   (iii) the substrate;    -   (c) contacting the patterned template surface with the        substrate; and    -   (d) treating the liquid material to form a pattern on the        substrate.

In some embodiments, the patterned template includes a solventresistant, low surface energy polymeric material derived from castinglow viscosity liquid materials onto a master template and then curingthe low viscosity liquid materials to generate a patterned template. Insome embodiments, the patterned template includes a solvent resistantelastomeric material.

In some embodiments, at least one of the patterned template andsubstrate includes a material selected from the group including aperfluoropolyether material, a fluoroolefin material, an acrylatematerial, a silicone material, a styrenic material, a fluorinatedthermoplastic elastomer (TPE), a triazine fluoropolymer, aperfluorocyclobutyl material, a fluorinated epoxy resin, and afluorinated monomer or fluorinated oligomer that can be polymerized orcrosslinked by a metathesis polymerization reaction.

In some embodiments, the perfluoropolyether material includes a backbonestructure selected from the group including:

wherein X is present or absent, and when present includes an endcappinggroup.

In some embodiments, the fluoroolefin material is selected from thegroup including:

wherein CSM includes a cure site monomer.

In some embodiments, the fluoroolefin material is made from monomerswhich include tetrafluoroethylene, vinylidene fluoride,hexafluoropropylene, 2,2-bis(trifluoromethyl)4,5-difluoro-1,3-dioxole, afunctional fluoroolefin, functional acrylic monomer, and a functionalmethacrylic monomer.

In some embodiments, the silicone material includes a fluoroalkylfunctionalized polydimethylsiloxane (PDMS) having the followingstructure:

wherein:

R is selected from the group including an acrylate, a methacrylate, anda vinyl group; and

Rf includes a fluoroalkyl chain.

In some embodiments, the styrenic material includes a fluorinatedstyrene monomer selected from the group including:

wherein Rf includes a fluoroalkyl chain.

In some embodiments, the acrylate material includes a fluorinatedacrylate or a fluorinated methacrylate having the following structure:

wherein:

R is selected from the group including H, alkyl, substituted alkyl,aryl, and substituted aryl; and

Rf includes a fluoroalkyl chain.

In some embodiments, the triazine fluoropolymer includes a fluorinatedmonomer.

In some embodiments, the fluorinated monomer or fluorinated oligomerthat can be polymerized or crosslinked by a metathesis polymerizationreaction includes a functionalized olefin. In some embodiments, thefunctionalized olefin includes a functionalized cyclic olefin.

In some embodiments, at least one of the patterned template and thesubstrate has a surface energy lower than 18 mN/m. In some embodiments,at least one of the patterned template and the substrate has a surfaceenergy lower than 15 mN/m. According to a further embodiment thepatterned template and/or the substrate has a surface energy betweenabout 10 mN/m and about 20 mN/m. According to some embodiments, thepatterned template and/or the substrate has a low surface energy ofbetween about 12 mN/m and about 15 mN/m.

In some embodiments, the substrate is selected from the group includinga polymer material, an inorganic material, a silicon material, a quartzmaterial, a glass material, and surface treated variants thereof. Insome embodiments, the substrate is selected from one of an electronicdevice in the process of being manufactured and a photonic device in theprocess of being manufactured. In some embodiments, the substrateincludes a patterned area.

In some embodiments, the plurality of recessed areas includes aplurality of cavities. In some embodiments, the plurality of cavitiesinclude a plurality of structural features. In some embodiments, theplurality of structural features has a dimension ranging from about 10microns to about 1 nanometer in size. In some embodiments, the pluralityof structural features has a dimension ranging from about 10 microns toabout 1 micron in size. In some embodiments, the plurality of structuralfeatures has a dimension ranging from about 1 micron to about 100 nm insize. In some embodiments, the plurality of structural features has adimension ranging from about 100 nm to about 1 nm in size. In someembodiments, the plurality of structural features has a dimension inboth the horizontal and vertical plane.

Referring now to FIGS. 39A-39F, one embodiment of a method for forming acomplex pattern on a substrate is illustrated. In the embodimentillustrated in FIG. 39, an imprint lithography technique is used to forma pattern on a substrate.

Referring now to FIG. 39A, a patterned master 3900 is provided.Patterned master 3900 includes a plurality of non-recessed surface 3920areas and a plurality of recesses 3930. In some embodiments, recesses3930 include one or more sub-recesses 3932. In some embodiments,recesses 3930 include a multiplicity of sub-recesses 3932. In someembodiments, patterned master 3900 includes an etched substrate, such asa silicon wafer, which is etched in the desired pattern to formpatterned master 3900.

Referring now to FIG. 39B, a flowable material 3901, for example, aliquid fluoropolymer composition, such as a PFPE-based precursor, ispoured onto patterned master 3900. In some embodiments, flowablematerial 3901 is treated by a treating process, for example exposure toUV light, thereby forming a treated material mold 3910 in the desiredpattern.

In one embodiment, illustrated in FIG. 39C, mold 3910 is removed frompatterned master 3900. In one embodiment, treated material mold 3910 isa cross-linked polymer. In one embodiment, treated material mold 3910 isan elastomer. In one embodiment, a force is applied to one or more ofmold 3910 or patterned master 3900 to separate mold 3910 from patternedmaster 3900. FIG. 39C illustrates one embodiment of mold 3910 andpatterned master 3900 wherein mold 3910 includes a plurality of recessesand sub-recesses which are mirror images of the plurality ofnon-recessed surface areas of patterned master 3900. In one embodimentof mold 3910 the plurality of non-recessed areas elastically deform tofacilitate removal of mold 3910 from master 3900. Mold 3910, in oneembodiment, is a useful patterned template for soft lithography andimprint lithography applications.

Referring now to FIG. 39D, a mold 3910 is provided. In some embodiments,mold 3910 includes a solvent resistant, low surface energy polymericmaterial, derived from casting low viscosity liquid materials onto amaster template and then curing the low viscosity liquid materials togenerate a patterned template as defined hereinabove. Mold 3910 furtherincludes a first patterned template surface 812 and a second templatesurface 814. The first patterned template surface 812 further includes aplurality of recesses 816 and subrecesses 3942. In one embodiment,multiple layers of subrecesses 3942 form sub-sub-recesses and so on. Insome embodiments, mold 3910 is derived from a solvent resistant, lowsurface energy polymeric material and is mounted on another material tofacilitate alignment of the mold or to facilitate continuous processing,such as a continuous process using a conveyor belt. In one embodiment,such continuous processing is useful in the fabrication of preciselyplaced structures on a surface, such as in the fabrication of a complexdevice or a semiconductor, electronic or photonic device.

Referring again to FIG. 39D, a substrate 3903 is provided. In someembodiments, substrate 3903 includes, without limitation, one or more ofa polymer material, an inorganic material, a silicon material, a quartzmaterial, a glass material, and surface treated variants thereof. Insome embodiments, at least one of mold 3910 and substrate 3903 has asurface energy lower than 18 mN/m. In some embodiments, at least one ofmold 3910 and substrate 3903 has a surface energy lower than 15 mN/m.According to a further embodiment the mold 3910 and/or the substrate3903 has a surface energy between about 10 mN/m and about 20 mN/m.According to some embodiments, the mold 3910 and/or the substrate 3903has a low surface energy of between about 12 mN/m and about 15 mN/m.

In some embodiments, as illustrated in FIG. 39D, mold 3910 and substrate3903 are positioned in a spaced relationship to each other such thatfirst patterned template surface 812 faces substrate surface 822 and agap 830 is created between first patterned template surface 812 and thesubstrate surface 822. This is merely one example of a predeterminedrelationship.

Referring again to FIG. 39D, a volume of liquid material 3902 isdisposed in the gap between first patterned template surface 812 andsubstrate surface 822. In some embodiments, the volume of liquidmaterial 3902 is disposed directly on a non-wetting agent (not shown),which is disposed on first patterned template surface 812.

Referring now to FIG. 39E, in some embodiments, mold 3910 is contactedwith the volume of liquid material 3902 (not shown in FIG. 39E). A forceF is applied to the mold 3910 thereby forcing the volume of liquidmaterial 3902 into the plurality of recesses 816 and sub-recesses. Insome embodiments, such as was illustrated in FIG. 8C, a portion of thevolume of liquid material 3902 remains between mold 3910 and substrate3903 surface after force F is applied.

Referring again to FIG. 39E, in some embodiments, the volume of liquidmaterial 3902 is treated by a treating process while force F is beingapplied to form a product 3904. In some embodiments, the treatingprocess includes, without limitation, one or more of a photochemicalprocess, a chemical process, combinations thereof, or the like.

Referring now to FIG. 39F, mold 3910 is removed from product 3904 toreveal a patterned product on substrate 3903 as shown in FIG. 39F. Insome embodiments, a residual, or “scum,” layer (not shown) of treatedliquid material remains on substrate 3903.

In some embodiments, the liquid material from which the particles willbe formed is selected from the group including a polymer, a solution, amonomer, a plurality of monomers, a polymerization initiator, apolymerization catalyst, an inorganic precursor, an organic material, anatural product, a metal precursor, a pharmaceutical agent, a tag, amagnetic material, a paramagnetic material, a superparamagneticmaterial, a ligand, a cell penetrating peptide, a porogen, a surfactant,a plurality of immiscible liquids, a solvent, a pharmaceutical agentwith a binder, and a charged species. In some embodiments, thepharmaceutical agent is selected from the group including a drug, apeptide, RNAi, and DNA. In some embodiments, the tag is selected fromthe group including a fluorescence tag, a radiolabeled tag, and acontrast agent. In some embodiments, the ligand includes a celltargeting peptide.

Representative superparamagnetic or paramagnetic materials include butare not limited to Fe₂O₃, Fe₃O₄, FePt, Co, MnFe₂O₄, CoFe₂O₄, CuFe₂O₄,NiFe₂O₄ and ZnS doped with Mn for magneto-optical applications, CdSe foroptical applications, and borates for boron neutron capture treatment.

In some embodiments, the liquid material is selected from one of aresist polymer and a low-k dielectric. In some embodiments, the liquidmaterial includes a non-wetting agent.

In some embodiments, the disposing of the volume of liquid material isregulated by a spreading process. In some embodiments, the spreadingprocess includes:

-   -   (a) disposing a first volume of liquid material on the patterned        template to form a layer of liquid material on the patterned        template; and    -   (b) drawing an implement across the layer of liquid material to:        -   (i) remove a second volume of liquid material from the layer            of liquid material on the patterned template; and        -   (ii) leave a third volume of liquid material on the            patterned template.            In some embodiments, the contacting of the first template            surface with the substrate eliminates essentially all of the            disposed volume of liquid material.

In some embodiments, the treating of the liquid includes, withoutlimitation, one or more of a thermal process, a photochemical process, achemical process, an evaporative process, a phase change, an oxidativeprocess, a reductive process, combinations thereof, or the like.

In some embodiments, the method includes a batch process. In someembodiments, the batch process is selected from one of a semi-batchprocess and a continuous batch process.

In some embodiments, the presently disclosed subject matter describes apatterned substrate formed by the presently disclosed methods.

IX. Imprint Lithography Free of a Residual “Scum Layer”

A characteristic of imprint lithography that has restrained its fullpotential is the formation of a “scum layer” once the liquid material,e.g., a resin, is patterned. The “scum layer” includes residual liquidmaterial that remains between the stamp and the substrate. In someembodiments, the presently disclosed subject matter provides a processfor generating patterns essentially free of a scum layer.

Referring now to FIGS. 9A-9E, in some embodiments, a method for forminga pattern on a substrate is provided, wherein the pattern is essentiallyfree of a scum layer. Referring now to FIG. 9A, a patterned template 910is provided. Patterned template 910 further includes a first patternedtemplate surface 912 and a second template surface 914. The firstpatterned template surface 912 further includes a plurality of recesses916. In some embodiments, a non-wetting agent 960 is disposed on thefirst patterned template surface 912.

Referring again to FIG. 9A, a substrate 920 is provided. Substrate 920includes a substrate surface 922. In some embodiments, a non-wettingagent 960 is disposed on substrate surface 920.

In some embodiments, as illustrated in FIG. 9A, patterned template 910and substrate 920 are positioned in a spaced relationship to each othersuch that first patterned template surface 912 faces substrate surface922 and a gap 930 is created between first patterned template surface912 and substrate surface 922.

Referring now to FIG. 9B, a volume of liquid material 940 is disposed inthe gap 930 between first patterned template surface 912 and substratesurface 922. In some embodiments, the volume of liquid material 940 isdisposed directly on first patterned template surface 912. In someembodiments, the volume of liquid material 940 is disposed directly onnon-wetting agent 960, which is disposed on first patterned templatesurface 912. In some embodiments, the volume of liquid material 940 isdisposed directly on substrate surface 920. In some embodiments, thevolume of liquid material 940 is disposed directly on non-wetting agent960, which is disposed on substrate surface 920.

Referring now to FIG. 9C, in some embodiments, first patterned templatesurface 912 is contacted with the volume of liquid material 940. A forceF_(a) is applied to second template surface 914 thereby forcing thevolume of liquid material 940 into the plurality of recesses 916. Incontrast with the embodiment illustrated in FIG. 8, a portion of thevolume of liquid material 940 is forced out of gap 930 by force F_(a)when force F_(a) is applied.

Referring again to FIG. 9C, in some embodiments, the volume of liquidmaterial 940 is treated by a treating process T_(r) while force F_(a) isbeing applied to form a treated liquid material 942.

Referring now to FIG. 9D, a force F_(r) is applied to patterned template910 to remove patterned template 910 from treated liquid material 942 toreveal a pattern 950 on substrate 920 as shown in FIG. 9E. In thisembodiment, substrate 920 is essentially free of a residual, or “scum,”layer of treated liquid material 942.

In some embodiments, at least one of the template surface and substrateincludes a functionalized surface element. In some embodiments, thefunctionalized surface element is functionalized with a non-wettingmaterial. In some embodiments, the non-wetting material includesfunctional groups that bind to the liquid material. In some embodiments,the non-wetting material is a trichloro silane, a trialkoxy silane, atrichloro silane including non-wetting and reactive functional groups, atrialkoxy silane including non-wetting and reactive functional groups,and/or mixtures thereof.

In some embodiments, the point of contact between the two surfaceelements is free of liquid material. In some embodiments, the point ofcontact between the two surface elements includes residual liquidmaterial. In some embodiments, the height of the residual liquidmaterial is less than 30% of the height of the structure. In someembodiments, the height of the residual liquid material is less than 20%of the height of the structure. In some embodiments, the height of theresidual liquid material is less than 10% of the height of thestructure. In some embodiments, the height of the residual liquidmaterial is less than 5% of the height of the structure. In someembodiments, the volume of liquid material is less than the volume ofthe patterned template. In some embodiments, substantially all of thevolume of liquid material is confined to the patterned template of atleast one of the surface elements. In some embodiments, having the pointof contact between the two surface elements free of liquid materialretards slippage between the two surface elements.

X. Solvent-Assisted Micro-molding (SAMIM)

In some embodiments, the presently disclosed subject matter describes asolvent-assisted micro-molding (SAMIM) method for forming a pattern on asubstrate.

Referring now to FIG. 10A, a patterned template 1010 is provided.Patterned template 1010 further includes a first patterned templatesurface 1012 and a second template surface 1014. The first patternedtemplate surface 1012 further includes a plurality of recesses 1016.

Referring again to FIG. 10A, a substrate 1020 is provided. Substrate1020 includes a substrate surface 1022. In some embodiments, a polymericmaterial 1070 is disposed on substrate surface 1022. In someembodiments, polymeric material 1070 includes a resist polymer.

Referring again to FIG. 10A, patterned template 1010 and substrate 1020are positioned in a spaced relationship to each other such that firstpatterned template surface 1012 faces substrate surface 1022 and a gap1030 is created between first patterned template surface 1012 andsubstrate surface 1022. As shown in FIG. 10A, a solvent S is disposedwithin gap 1030, such that solvent S contacts polymeric material 1070forming a swollen polymeric material 1072.

Referring now to FIGS. 10B and 10C, first patterned template surface1012 is contacted with swollen polymeric material 1072. A force F_(a) isapplied to second template surface 1014 thereby forcing a portion ofswollen polymeric material 1072 into the plurality of recesses 1016 andleaving a portion of swollen polymeric material 1072 between firstpatterned template surface 1012 and substrate surface 1020. The swollenpolymeric material 1072 is then treated by a treating process T_(r)while under pressure.

Referring now to FIG. 10D, a force F_(r) is applied to patternedtemplate 1010 to remove patterned template 1010 from treated swollenpolymeric material 1072 to reveal a polymeric pattern 1074 on substrate1020 as shown in FIG. 10E.

XI. Removing the Patterned Structure from the Patterned Template and/orSubstrate

In some embodiments, the patterned structure (e.g., a patterned micro-or nanostructure) is removed from at least one of the patterned templateand/or the substrate. This can be accomplished by a number ofapproaches, including but not limited to applying the surface elementcontaining the patterned structure to a surface that has an affinity forthe patterned structure; applying the surface element containing thepatterned structure to a material that when hardened has a chemicaland/or physical interaction with the patterned structure; deforming thesurface element containing the patterned structure such that thepatterned structure is released from the surface element; swelling thesurface element containing the patterned structure with a first solventto extrude the patterned structure; and washing the surface elementcontaining the patterned structure with a second solvent that has anaffinity for the patterned structure.

In some embodiments, the surface that has an affinity for the particlesincludes an adhesive or sticky surface (e.g. carbohydrates, epoxies,waxes, polyvinyl alcohol, polyvinyl pyrrolidone, polybutyl acrylate,polycyano acrylates, polymethyl methacrylate). In some embodiments, theliquid is water that is cooled to form ice. In some embodiments, thewater is cooled to a temperature below the Tm of water but above the Tgof the particle. In some embodiments the water is cooled to atemperature below the Tg of the particles but above the Tg of the moldor substrate. In some embodiments, the water is cooled to a temperaturebelow the Tg of the mold or substrate.

In some embodiments, the first solvent includes supercritical fluidcarbon dioxide. In some embodiments, the first solvent includes water.In some embodiments, the first solvent includes an aqueous solutionincluding water and a detergent. In embodiments, the deforming thesurface element is performed by applying a mechanical force to thesurface element. In some embodiments, the method of removing thepatterned structure further includes a sonication method.

XII. Method of Fabricating Molecules and for Delivering a TherapeuticAgent to a Target

In some embodiments, the presently disclosed subject matter describesmethods, processes, and products by processes, for fabricating deliverymolecules, for use in drug discovery and drug therapies. In someembodiments, the method or process for fabricating a delivery moleculeincludes a combinatorial method or process. In some embodiments, themethod for fabricating molecules includes a non-wetting imprintlithography method.

XII.A. Method of Fabricating Molecules

In some embodiments, the non-wetting imprint lithography method of thepresently disclosed subject matter is used to generate a surface derivedfrom or including a solvent resistant, low surface energy polymericmaterial. The surface is derived from casting low viscosity liquidmaterials onto a master template and then curing the low viscosityliquid materials to generate a patterned template, as described herein.In some embodiments, the surface includes a solvent resistantelastomeric material.

In some embodiments, the non-wetting imprint lithography method is usedto generate isolated structures. In some embodiments, the isolatedstructures include isolated micro-structures. In some embodiments, theisolated structures include isolated nano-structures. In someembodiments, the isolated structures include a biodegradable material.In some embodiments, the isolated structures include a hydrophilicmaterial. In some embodiments, the isolated structures include ahydrophobic material. In some embodiments, the isolated structuresinclude a particular shape. In another embodiment, the isolatedstructures include or are configured to hold include “cargo.” Accordingto one embodiment, the cargo held by the isolated structure can includean element, a molecule, a chemical substance, an agent, a drug, abiologic, a protein, DNA, RNA, a diagnostic, a therapeutic, a cancertreatment, a viral treatment, a bacterial treatment, a fungal treatment,an auto-immune treatment, combinations thereof, or the like. Accordingto an alternative embodiment, the cargo protrudes from the surface ofthe isolated structure, thereby functionalizing the isolated structure.According to yet another embodiment, the cargo is completely containedwithin the isolated particle such that the cargo is stealthed orsheltered from an environment to which the isolated structure can besubjected. According to yet another embodiment, the cargo is containedsubstantially on the surface of the isolated structure. In a furtherembodiment, the cargo is associated with the isolated structure in acombination of one of the above techniques, or the like.

According to another embodiment, the cargo is attached to the isolatedstructure by chemical binding or physical constraint. In someembodiments, the chemical binding includes, but is not limited to,covalent binding, ionic bonding, other intra- and inter-molecularforces, hydrogen bonding, van der Waals forces, combinations thereof,and the like.

In some embodiments, the non-wetting imprint lithography method furtherincludes adding molecular modules, fragments, or domains to the solutionto be molded. In some embodiments, the molecular modules, fragments, ordomains impart functionality to the isolated structures. In someembodiments, the functionality imparted to the isolated structureincludes a therapeutic functionality.

In some embodiments, a therapeutic agent, such as a drug, a biologic,combinations thereof, and the like, is incorporated into the isolatedstructure. In some embodiments, the physiologically active drug istethered to a linker to facilitate its incorporation into the isolatedstructure. In some embodiments, the domain of an enzyme or a catalyst isadded to the isolated structure. In some embodiments, a ligand or anoligopeptide is added to the isolated structure. In some embodiments,the oligopeptide is functional. In some embodiments, the functionaloligopeptide includes a cell targeting peptide. In some embodiments, thefunctional oligopeptide includes a cell penetrating peptide. In someembodiments an antibody or functional fragment thereof is added to theisolated structure.

In some embodiments, a binder is added to the isolated structure. Insome embodiments, the isolated structure including the binder is used tofabricate identical structures. In some embodiments, the isolatedstructure including the binder is used to fabricate structures of avarying structure. In some embodiments, the structures of a varyingstructure are used to explore the efficacy of a molecule as atherapeutic agent. In some embodiments, the shape of the isolatedstructure mimics a biological agent. In some embodiments, the methodfurther includes a method for drug discovery.

XII.B. Method of Delivering a Therapeutic Agent to a Target

In some embodiments, a method of delivering a therapeutic agent to atarget is disclosed, the method including: providing a particle producedas described herein; admixing the therapeutic agent with the particle;and delivering the particle including the therapeutic agent to thetarget.

In some embodiments, the therapeutic agent includes a drug. In someembodiments, the therapeutic agent includes genetic material. In someembodiments, the genetic material includes, without limitation, one ormore of a non-viral gene vector, DNA, RNA, RNAi, a viral particle,combinations thereof, or the like.

In some embodiments, the particle has a diameter of less than 100microns. In some embodiments, the particle has a diameter of less than10 microns. In some embodiments, the particle has a diameter of lessthan 1 micron. In some embodiments, the particle has a diameter of lessthan 100 nm. In some embodiments, the particle has a diameter of lessthan 10 nm.

In some embodiments, the particle includes a biodegradable polymer. Abiodegradable polymer is defined as a polymer that undergoes a reductionin molecular weight upon either a change in biological condition orexposure to a biological agent. In some embodiments, the biodegradablepolymer includes, without limitation, one or more of a polyester, apolyanhydride, a polyamide, a phosphorous-based polymer, apoly(cyanoacrylate), a polyurethane, a polyorthoester, apolydihydropyran, a polyacetal, combinations thereof, or the like. Insome embodiments, the polymer is modified to be a biodegradable polymer(e.g. a poly(ethylene glycol) that is functionalized with a disulfidegroup). In some embodiments, the polyester includes, without limitation,one or more of polylactic acid, polyglycolic acid,poly(hydroxybutyrate), poly(ε-caprolactone), poly(β-malic acid),poly(dioxanones), combinations thereof, or the like. In someembodiments, the polyanhydride includes, without limitation, one or moreof poly(sebacic acid), poly(adipic acid), poly(terpthalic acid),combinations thereof, or the like. In some embodiments, the polyamideincludes, without limitation, one or more of a poly(imino carbonate), apolyaminoacid, combinations thereof, or the like. In some embodiments,the phosphorous-based polymer includes, without limitation, one or moreof polyphosphates, polyphosphonates, polyphosphazenes, combinationsthereof, or the like. In some embodiments, the polymer is responsive tostimuli, such as pH, radiation, oxidation, reduction, ionic strength,temperature, and alternating magnetic or electric fields.

Responses to such stimuli can include swelling, bond cleavage, heating,combinations thereof, or the like, which can facilitate release of theisolated structures cargo, degradation of the isolated structure itself,combinations thereof, and the like.

In some embodiments, the presently disclosed subject matter describesmagneto containing particles for applications in hyperthermia therapy,cancer and gene therapy, drug delivery, magnetic resonance imagingcontrast agents, vaccine adjuvants, memory devices, spintronics,combinations thereof, and the like.

Without being bound to any one particular theory, the magneto containingparticles, e.g., a magnetic nanoparticle, produce heat by the process ofhyperthermia (between 41 and 46° C.) or thermo ablation (greater than46° C.), i.e., the controlled heating of the nanoparticles upon exposureto an AC-magnetic field. The heat is used to (i) induce a phase changein the polymer component (for example melt and release an encapsulatedmaterial) and/or (ii) hyperthermia treatment of specific cells and/or(iii) increase the effectiveness of the encapsulated material. Thetriggering mechanism of the magnetic nanoparticles via electromagneticheating enhance the (iv) degradation rate of the particulate; (v) caninduce swelling; and/or (vi) induce dissolution/phase change that canlead to a greater surface area, which can be beneficial when treating avariety of diseases.

In some embodiments, the presently disclosed subject matter describes analternative therapeutic agent delivery method, which utilizes“non-wetting” imprint lithography to fabricate monodisperse magneticnanoparticles for use in a drug delivery system. Such particles can beused for: (1) hyperthermia treatment of cancer cells; (2) MRI contrastagents; (3) guided delivery of the particle; and (4) triggereddegradation of the drug delivery vector.

In some embodiments, the therapeutic agent delivery system includes abiocompatible material and a magnetic nanoparticle. In some embodiments,the biocompatible material has a melting point below 100° C. In someembodiments, the biocompatible material includes, without limitation,one or more of a polylactide, a polyglycolide, a hydroxypropylcellulose,a wax, combinations thereof, or the like.

In some embodiments, once the magnetic nanoparticle is delivered to thetarget or is in close proximity to the target, the magnetic nanoparticleis exposed to an AC-magnetic field. The exposure to the AC-magneticfield causes the magnetic nanoparticle to undergo a controlled heating.Without being bound to any one particular theory, the controlled heatingis a result of a thermo ablation process. In some embodiments, the heatis used to induce a phase change in the polymer component of thenanoparticle. In some embodiments, the phase change includes a meltingprocess. In some embodiments, the phase change results in the release ofan encapsulated material. In some embodiments, the release of anencapsulated material includes a controlled release. In someembodiments, the controlled release of the encapsulated material resultsin a concentrated dosing of the therapeutic agent. In some embodiments,the heating results in the hyperthermic treatment of the target, e.g.,specific cells. In some embodiments, the heating results in an increasein the effectiveness of the encapsulated material. In some embodiments,the triggering mechanism of the magnetic nanoparticles induced by theelectromagnetic heating enhances the degradation rate of the particleand can induce swelling and/or a dissolution/phase change that can leadto a greater surface area which can be beneficial when treating avariety of diseases.

The presently described magnetic containing materials also lendthemselves to other applications. The magneto-particles can be assembledinto well-defined arrays driven by their shape, functionalization of thesurface and/or exposure to a magnetic field for investigations of andnot limited to magnetic assay devices, memory devices, spintronicapplications, and separations of solutions.

Thus, the presently disclosed subject matter provides a method fordelivering a therapeutic agent to a target, the method including:

-   -   (a) providing a particle prepared by the presently disclosed        methods;    -   (b) admixing the therapeutic agent with the particle; and    -   (c) delivering the particle including the therapeutic agent to        the target.

In some embodiments, the method includes exposing the particle to analternating magnetic field once the particle is delivered to the target.In some embodiments, the exposing of the particle to an alternatingmagnetic field causes the particle to produce heat through one of ahypothermia process, a thermo ablation process, combinations thereof, orthe like.

In some embodiments, the heat produced by the particle induces one of aphase change in the polymer component of the particle and a hyperthermictreatment of the target. In some embodiments, the phase change in thepolymer component of the particle includes a change from a solid phaseto a liquid phase. In some embodiments, the phase change from a solidphase to a liquid phase causes the therapeutic agent to be released fromthe particle. In some embodiments, the release of the therapeutic agentfrom the particle includes a controlled release.

In some embodiments, the target includes, without limitation, one ormore of a cell-targeting peptide, a cell-penetrating peptide, anintegrin receptor peptide (GRGDSP), a melanocyte stimulating hormone, avasoactive intestional peptide, an anti-Her2 mouse antibody, a vitamin,combinations thereof, or the like.

In one embodiment, the presently disclosed subject matter provides amethod for modifying a particle surface. In one embodiment the method ofmodifying a particle surface includes: (a) providing particles in or onat least one of: (i) a patterned template; and (ii) a substrate; (b)disposing a solution containing a modifying group in or on at least oneof: (i) the patterned template; and (ii) the substrate; and (c) removingexcess unreacted modifying groups.

In one embodiment of the method for modifying a particle, the modifyinggroup chemically attaches to the particle through a linking group. Inanother embodiment of the method for modifying a particle, the linkergroup includes, without limitation, one or more of sulfides, amines,carboxylic acids, acid chlorides, alcohols, alkenes, alkyl halides,isocyanates, combinations thereof, or the like. In another embodiment,the method of modifying the particles includes a modifying agent thatincludes, without limitation, one or more of dyes, fluorescence tags,radiolabeled tags, contrast agents, ligands, peptides, pharmaceuticalagents, proteins, DNA, RNA, siRNA, combinations thereof, or the like.

With respect to the methods of the presently disclosed subject matter,any animal subject can be treated. The term “subject” as used hereinrefers to any vertebrate species. The methods of the presently claimedsubject matter are particularly useful in the diagnosis of warm-bloodedvertebrates. Thus, the presently claimed subject matter concernsmammals. In some embodiments provided is the diagnosis and/or treatmentof mammals such as humans, as well as those mammals of importance due tobeing endangered (such as Siberian tigers), of economical importance(animals raised on farms for consumption by humans) and/or socialimportance (animals kept as pets or in zoos) to humans, for instance,carnivores other than humans (such as cats and dogs), swine (pigs, hogs,and wild boars), ruminants (such as cattle, oxen, sheep, giraffes, deer,goats, bison, and camels), and horses. Also provided is the diagnosisand/or treatment of livestock, including, but not limited todomesticated swine (pigs and hogs), ruminants, horses, poultry, and thelike.

The following references are incorporated herein by reference in theirentirety. Published International PCT Application No. WO2004081666 toDeSimone et al.; U.S. Pat. No. 6,528,080 to Dunn et al.; U.S. Pat. No.6,592,579 to Arndt et al., Published International PCT Application No.WO0066192 to Jordan; Hilger, I. et al., Radiology 570-575 (2001);Mornet, S. et al., J. Mat. Chem., 2161-2175 (2004); Berry, C. C. et al.,J. Phys. D: Applied Physics 36, R198-R206 (2003); Babincova, M. et al.,Bioelectrochemistry 55, 17-19 (2002); Wolf, S. A. et al., Science 16,1488-1495 (2001); and Sun, S. et al., Science 287, 1989-1992 (2000);U.S. Pat. No. 6,159,443 to Hallahan; and Published PCT Application No.WO 03/066066 to Hallahan et al.

XIII. Method of Patterning Natural and Synthetic Structures

In some embodiments, the presently disclosed subject matter describesmethods and processes, and products by processes, for generatingsurfaces and molds from natural structures, single molecules, orself-assembled structures. Accordingly, in some embodiments, thepresently disclosed subject matter describes a method of patterning anatural structure, single molecule, and/or a self-assembled structure.In some embodiments, the method further includes replicating the naturalstructure, single molecule, and/or a self-assembled structure. In someembodiments, the method further includes replicating the functionalityof the natural structure, single molecule, and/or a self-assembledstructure.

More particularly, in some embodiments, the method further includestaking the impression or mold of a natural structure, single molecule,and/or a self-assembled structure. In some embodiments, the impressionor mold is taken with a low surface energy polymeric precursor. In someembodiments, the low surface energy polymeric precursor includes aperfluoropolyether (PFPE) functionally terminated diacrylate. In someembodiments, the natural structure, single molecule, and/orself-assembled structure includes, without limitation, one or more ofenzymes, viruses, antibodies, micelles, tissue surfaces, combinationsthereof, or the like.

In some embodiments, the impression or mold is used to replicate thefeatures of the natural structure, single molecule, and/or aself-assembled structure into an isolated object or a surface. In someembodiments, a non-wetting imprint lithography method is used to impartthe features into a molded part or surface. In some embodiments, themolded part or surface produced by this process can be used in manyapplications, including, but not limited to, drug delivery, medicaldevices, coatings, catalysts, or mimics of the natural structures fromwhich they are derived. In some embodiments, the natural structureincludes biological tissue. In some embodiments, the biological tissueincludes tissue from a bodily organ, such as a heart. In someembodiments, the biological tissue includes vessels and bone. In someembodiments, the biological tissue includes tendon or cartilage. Forexample, in some embodiments, the presently disclosed subject matter canbe used to pattern surfaces for tendon and cartilage repair. Such repairtypically requires the use of collagen tissue, which comes from cadaversand must be machined for use as replacements. Most of these replacementsfail because one cannot lay down the primary pattern that is requiredfor replacement. The soft lithographic methods described hereinalleviate this problem.

In some embodiments, the presently disclosed subject matter can beapplied to tissue regeneration using stem cells. Almost all stem cellapproaches known in the art require molecular patterns for the cells toseed and then grow, thereby taking the shape of an organ, such as aliver, a kidney, or the like. In some embodiments, the molecularscaffold is cast and used as crystals to seed an organ in a form oftransplant therapy. In some embodiments, the stem cell andnano-substrate is seeded into a dying tissue, e.g., liver tissue, topromote growth and tissue regeneration. In some embodiments, thematerial to be replicated in the mold includes a material that issimilar to or the same as the material that was originally molded. Insome embodiments, the material to be replicated in the mold includes amaterial that is different from and/or has different properties than thematerial that was originally molded. This approach could play animportant role in addressing the organ transplant shortage.

In some embodiments, the presently disclosed subject matter is used totake the impression of one of an enzyme, a bacterium, and a virus. Insome embodiments, the enzyme, bacterium, or virus is then replicatedinto a discrete object or onto a surface that has the shape reminiscentof that particular enzyme, bacterium, or virus replicated into it. Insome embodiments, the mold itself is replicated on a surface, whereinthe surface-attached replicated mold acts as a receptor site for anenzyme, bacterium, or virus particle. In some embodiments, thereplicated mold is useful as a catalyst, a diagnostic sensor, atherapeutic agent, a vaccine, combinations thereof, and the like. Insome embodiments, the surface-attached replicated mold is used tofacilitate the discovery of new therapeutic agents.

In some embodiments, the macromolecular, e.g., enzyme, bacterial, orviral, molded “mimics” serve as non-self-replicating entities that havethe same surface topography as the original macromolecule, bacterium, orvirus. In some embodiments, the molded mimics are used to createbiological responses, e.g., an allergic response, to their presence,thereby creating antibodies or activating receptors. In someembodiments, the molded mimics function as a vaccine. In someembodiments, the efficacy of the biologically-active shape of the moldedmimics is enhanced by a surface modification technique.

XIII.A. Molecular Imprinting

According to some embodiments, the materials and methods of thepresently disclosed subject matter can be used with molecular imprintingtechniques to form polymers with recognition cites. Drug research anddevelopment often requires the analysis of highly specific and sensitivechemical and/or biologic agents collectively called “recognitionagents.” Natural recognition agents, such as for example, enzymes,proteins, drug candidates, biomolecules, herbicides, amino acids,derivatives of amino acids, peptides, nucleotides, nucleotide bases,combinations thereof, and the like, tend to be very specific andsensitive as well as being labile and have a low density of bindingsites. Because of the delicacy of natural recognition agents, artificialrecognition agents are more stable and have become popular researchtools. Molecular imprinting has emerged in recent years as a highlyaccepted tool for the development of artificial recognition agents.

Imprinting of molecules occurs by the polymerization of functional andcross-linking monomers in the presence of a template molecule. First, atemplate molecule, such as, by way of example but not limitation, anenzyme, a protein, a drug candidate, a biomolecule, a herbicide, anamino acid, a derivative of an amino acid, a peptide, nucleotides,nucleotide bases, a virus, combinations thereof, and the like isintroduced to a liquid polymer solution. In some embodiments, the liquidpolymer solution is a liquid polymer of the presently disclosed subjectmatter and includes functional and cross-linked monomers. The functionaland cross-linked monomers are allowed to establish bond formations andother chemical and physical associations and orientations with thetemplate in the polymer. In some embodiments, a functional monomerincludes two functional groups. At one end of the monomer the monomer isconfigured to interact with the template, for example throughnoncovalent interactions (i.e., hydrogen bonding, van der Waals forces,or hydrophobic interactions). The other end of the monomer, i.e., theend that is not interacting with the template, includes a group that isable to bind with the polymer. During polymerization, the monomers arelocked in position around the template, for example with covalentbinding, and remain in such a position after the template is removed.

After polymerization or curing the template is removed from the polymer.The template can be removed by dissolving the template in a solvent insome embodiments. The resultant imprint of the template has a steric(size and shape) and chemical (spatial arrangements or complementaryfunctionality) memory of the template. After polymerization and removalof the template, the functional groups of the polymers molecular imprintcan then bind a target provided that the binding sites of the imprintand the target molecule complement each other in size, shape, andchemical functionality. This process provides a material with a highstability against physicochemical perturbations that has specificitytoward a target molecule and, as such, the material can be used in highthroughput assays and in conjunction with physical and chemicalparameters that a natural recognition agent cannot withstand.

According to some embodiments, applications of molecular imprintinginclude, but are not limited to, purification, separation, screening ofbioactive molecules, sensors, catalysis, chromatographic separation,drug screening, chemosensors, catalysis, biodefense, immunoassays,combinations thereof, and the like.

Useful applications and experimentations of molecular imprinting thatcan be used in combination with the materials and methods of thepresently disclosed subject matter can be found in: Vivek BabuKandimalla, Hunagxian Ju, Molecular Imprinting: A Dynamic Technique forDiverse Applications in Analytical Chemistry, Anal. Bioanal. Chem.(2004) 380: 587-605, and the references cited therein, which are allhereby incorporated by reference in their entirety herein.

XIII.B. Artificial Functional Molecules

According to some embodiments of the presently disclosed subject matter,following the formation of a molecular imprint of a template molecule,as described herein, the molecular imprint can then be used as a moldand receive the materials and methods of the presently disclosed subjectmatter to form, for example, an artificial functional molecule. Afterforming the functionalized molecular imprint mold in the polymermaterial, a polymer precursor solution including, but not limited to,functional and cross-linked monomers, can be applied to thefunctionalized imprint mold in accord with the materials and methodsdisclosed herein to form an artificial functional molecule. Duringmolding of the artificial functional molecule, the functionalizedmonomers in the polymer precursor will align with the functionalizedparts of the imprint mold such that the artificial functional moleculewill posses a steric (size and shape) and chemical (spatial arrangementsor complementary functionality) memory of the imprint mold. Theartificial functional molecule, which is the steric and chemical memoryof the imprint mold, has similar chemical and physical properties to theoriginal template molecule and can trigger membrane channels; bind toreceptors; enter cells; interact with proteins and enzymes; triggerimmune responses; trigger physiological responses; trigger release ofbioregulatory agents such as, for example, hormones, “feel good”molecules, neurotransmitters, and the like; inhibit responses; triggerregulatory functions; combinations thereof; and the like.

According to other embodiments, molecular imprints and artificialfunctional molecules of the presently disclosed subject matter can beused in conjunction with particles of the presently disclosed subjectmatter, as disclosed herein, that have drugs, biologics, or other agentsfor analysis associated with the particle. Accordingly, the particleswith drugs, biologics, or other agents can be analyzed for interactionand/or binding with the artificial functional molecule particles and/ormolecular imprint, thereby, making a complete analysis system havinghigh stability against physicochemical perturbations and, as such, thematerials can be used in high throughput assays and in conjunction withphysical and chemical parameters that natural recognition agents can notwithstand. Further, the presently disclosed analysis systems made of thematerials and methods of the presently disclosed subject matter areeconomical to manufacture, increase throughput of drug and biomoleculeresearch and development, and the like.

Referring now to FIG. 44, an embodiment of forming an artificialfunctional molecule includes creating a molecular imprinting such asshown in FIG. 44A. A substrate material 4410, such as liquidperfluoropolyether, contains functional monomers 4412 and 4414.Substrate material 4410 is imprinted with template molecules 4420 havingspecific steric and chemical groupings 4418 associated therewith.Template molecules 4420 form imprint wells 4416 in substrate material4410. Substrate material 4410 is then cured, for example by photocuring,thermal curing, combinations thereof, or the like as described herein.

Next, in FIG. 44B, template molecules 4420 are removed, dissociated, ordissolved from association with substrate material 4410. Before curingof substrate material 4410, however, functional monomers 4412 and 4414of substrate material 4410 associate with their negative or mirror imagein template molecules 4420 and during polymerization the functionalmonomers become locked in position. Thereby, a molecular imprint 4430that is the steric and chemical mirror image of the template molecule4420 is formed in the substrate material.

Next, an artificial functional molecule 4440 is formed in molecularimprint 4430. According to an embodiment, the materials and methods ofthe presently disclosed subject matter are utilized, as describedelsewhere herein, to make particles that mimic, both stericly andchemically template molecule 4420 that made imprint 4430. According toone embodiment, liquid PFPE is prepared and mixed with functionalmonomers 4444 and the mixture is introduced into molecular imprintcavity 4442 in substrate 4410. Functional monomers 4444 in the PFPEassociate with their mirror image functional monomer 4412 and 4414 lockinto place in substrate material 4410. The PFPE mixture is then curedsuch that artificial functional molecules 4440 are formed in imprintcavity 4442 and mimic template molecule 4420 both stericly andchemically. Artificial functional molecules 4444 are then removed fromthe substrate 4410 as described herein.

XIV. Method of Modifying the Surface of an Imprint Lithography Mold toImpart Surface Characteristics to Molded Products

In some embodiments, the presently disclosed subject matter describes amethod of modifying the surface of an imprint lithography mold. In someembodiments, the method further includes imparting surfacecharacteristics to a molded product. In some embodiments, the moldedproduct includes an isolated molded product. In some embodiments, theisolate molded product is formed using a non-wetting imprint lithographytechnique. In some embodiments, the molded product includes a contactlens, a medical device, and the like.

More particularly, the surface of a solvent resistant, low surfaceenergy polymeric material, or more particularly a PFPE mold is modifiedby a surface modification step, wherein the surface modification stepincludes, without limitation, one or more of plasma treatment, chemicaltreatment, the adsorption of molecules, combinations thereof, or thelike. In some embodiments, the molecules adsorbed during the surfacemodification step include, without limitation, one or more ofpolyelectrolytes, poly(vinylalcohol), alkylhalosilanes, ligands,combinations thereof, or the like. In some embodiments, the structures,particles, or objects obtained from the surface-treated molds can bemodified by the surface treatments in the mold. In some embodiments, themodification includes the pre-orientation of molecules or moieties withthe molecules including the molded products. In some embodiments, thepre-orientation of the molecules or moieties imparts certain propertiesto the molded products, including catalytic, wettable, adhesive,non-stick, interactive, or not interactive, when the molded product isplaced in another environment. In some embodiments, such properties areused to facilitate interactions with biological tissue or to preventinteraction with biological tissues. Applications of the presentlydisclosed subject matter include sensors, arrays, medical implants,medical diagnostics, disease detection, and separation media.

XV. Methods for Selectively Exposing the Surface of an Article to anAgent

Also disclosed herein is a method for selectively exposing the surfaceof an article to an agent. In some embodiments the method includes:

-   -   (a) shielding a first portion of the surface of the article with        a masking system, wherein the masking system includes a        elastomeric mask in conformal contact with the surface of the        article; and    -   (b) applying an agent to be patterned within the masking system        to a second portion of the surface of the article, while        preventing application of the agent to the first portion        shielded by the masking system.

In some embodiments, the elastomeric mask includes a plurality ofchannels. In some embodiments, each of the channels has across-sectional dimension of less than about 1 millimeter. In someembodiments, each of the channels has a cross-sectional dimension ofless than about 1 micron. In some embodiments, each of the channels hasa cross-sectional dimension of less than about 100 nm. In someembodiments, each of the channels has a cross-sectional dimension ofabout 1 nm. In some embodiments, the agent swells the elastomeric maskless than 25%.

In some embodiments, the agent includes an organic electroluminescentmaterial or a precursor thereof. In some embodiments, the method furtherincluding allowing the organic electroluminescent material to form fromthe agent at the second portion of the surface, and establishingelectrical communication between the organic electroluminescent materialand an electrical circuit.

In some embodiments, the agent includes a liquid or is carried in aliquid. In some embodiments, the agent includes the product of chemicalvapor deposition. In some embodiments, the agent includes a product ofdeposition from a gas phase. In some embodiments, the agent includes aproduct of e-beam deposition, evaporation, or sputtering. In someembodiments, the agent includes a product of electrochemical deposition.In some embodiments, the agent includes a product of electrolessdeposition. In some embodiments, the agent is applied from a fluidprecursor. In some embodiments, includes a solution or suspension of aninorganic compound. In some embodiments, the inorganic compound hardenson the second portion of the article surface.

In some embodiments, the fluid precursor includes a suspension ofparticles in a fluid carrier. In some embodiments, the method furtherincludes allowing the fluid carrier to dissipate thereby depositing theparticles at the first region of the article surface. In someembodiments, the fluid precursor includes a chemically active agent in afluid carrier. In some embodiments, the method further includes allowingthe fluid carrier to dissipate thereby depositing the chemically activeagent at the first region of the article surface.

In some embodiments, the chemically active agent includes a polymerprecursor. In some embodiments, the method further includes forming apolymeric article from the polymer precursor. In some embodiments, thechemically active agent includes an agent capable of promotingdeposition of a material. In some embodiments, the chemically activeagent includes an etchant. In some embodiments, the method furtherincludes allowing the second portion of the surface of the article to beetched. In some embodiments, the method further includes removing theelastomeric mask of the masking system from the first portion of thearticle surface while leaving the agent adhered to the second portion ofthe article surface.

XVI. Methods for Forming Engineered Membranes

The presently disclosed subject matter also describes a method forforming an engineered membrane. In some embodiments, a patternednon-wetting template is formed by contacting a first liquid material,such as a PFPE material, with a patterned substrate and treating thefirst liquid material, for example, by curing through exposure to UVlight to form a patterned non-wetting template. The patterned substrateincludes a plurality of recesses or cavities configured in a specificshape such that the patterned non-wetting template includes a pluralityof extruding features. The patterned non-wetting template is contactedwith a second liquid material, for example, a photocurable resin. Aforce is then applied to the patterned non-wetting template to displacean excess amount of second liquid material or “scum layer.” The secondliquid material is then treated, for example, by curing through exposureto UV light to form an interconnected structure including a plurality ofshape and size specific holes. The interconnected structure is thenremoved from the non-wetting template. In some embodiments, theinterconnected structure is used as a membrane for separations.

XVII. Methods for Inspecting Processes and Products by Processes

It will be important to inspect the objects/structures/particlesdescribed herein for accuracy of shape, placement and utility. Suchinspection can allow for corrective actions to be taken or for defectsto be removed or mitigated. The range of approaches and monitoringdevices useful for such inspections include: air gages, which usepneumatic pressure and flow to measure or sort dimensional attributes;balancing machines and systems, which dynamically measure and/or correctmachine or component balance; biological microscopes, which typicallyare used to study organisms and their vital processes; bore and IDgages, which are designed for internal diameter dimensional measurementor assessment; boroscopes, which are inspection tools with rigid orflexible optical tubes for interior inspection of holes, bores,cavities, and the like; calipers, which typically use a precise slidemovement for inside, outside, depth or step measurements, some of whichare used for comparing or transferring dimensions; CMM probes, which aretransducers that convert physical measurements into electrical signals,using various measuring systems within the probe structure; color andappearance instruments, which, for example, typically are used tomeasure the properties of paints and coatings including color, gloss,haze and transparency; color sensors, which register items by contrast,true color, or translucent index, and are based on one of the colormodels, most commonly the RGB model (red, green, blue); coordinatemeasuring machines, which are mechanical systems designed to move ameasuring probe to determine the coordinates of points on a work piecesurface; depth gages, which are used to measure of the depth of holes,cavities or other component features; digital/video microscopes, whichuse digital technology to display the magnified image; digital readouts,which are specialized displays for position and dimension readings frominspection gages and linear scales, or rotary encoders on machine tools;dimensional gages and instruments, which provide quantitativemeasurements of a product's or component's dimensional and formattributes such as wall thickness, depth, height, length, I.D., O.D.,taper or bore; dimensional and profile scanners, which gathertwo-dimensional or three-dimensional information about an object and areavailable in a wide variety of configurations and technologies; electronmicroscopes, which use a focused beam of electrons instead of light to“image” the specimen and gain information as to its structure andcomposition; fiberscopes, which are inspection tools with flexibleoptical tubes for interior inspection of holes, bores, and cavities;fixed gages, which are designed to access a specific attribute based oncomparative gaging, and include Angle Gages, Ball Gages, Center Gages,Drill Size Gages, Feeler Gages, Fillet Gages, Gear Tooth Gages, Gage orShim Stock, Pipe Gages, Radius Gages, Screw or Thread Pitch Gages, TaperGages, Tube Gages, U.S. Standard Gages (Sheet/Plate), Weld Gages andWire Gages; specialty/form gages, which are used to inspect parameterssuch as roundness, angularity, squareness, straightness, flatness,runout, taper and concentricity; gage blocks, which are manufactured toprecise gagemaker tolerance grades for calibrating, checking, andsetting fixed and comparative gages; height gages, which are used formeasuring the height of components or product features; indicators andcomparators, which measure where the linear movement of a precisionspindle or probe is amplified; inspection and gaging accessories, suchas layout and marking tolls, including hand tools, supplies andaccessories for dimensional measurement, marking, layout or othermachine shop applications such as scribes, transfer punches, dividers,and layout fluid; interferometers, which are used to measure distance interms of wavelength and to determine wavelengths of particular lightsources; laser micrometers, which measure extremely small distancesusing laser technology; levels, which are mechanical or electronic toolsthat measure the inclination of a surface relative to the earth'ssurface; machine alignment equipment, which is used to align rotating ormoving parts and machine components; magnifiers, which are inspectioninstruments that are used to magnify a product or part detail via a lenssystem; master and setting gages, which provide dimensional standardsfor calibrating other gages; measuring microscopes, which are used bytoolmakers for measuring the properties of tools, and often are used fordimensional measurement with lower magnifying powers to allow forbrighter, sharper images combined with a wide field of view;metallurgical microscopes, which are used for metallurgical inspection;micrometers, which are instruments for precision dimensional gagingincluding a ground spindle and anvil mounted in a C-shaped steel frame.Noncontact laser micrometers are also available; microscopes (alltypes), which are instruments that are capable of producing a magnifiedimage of a small object; optical fight microscopes, which use thevisible or near-visible portion of the electromagnetic spectrum; opticalcomparators, which are instruments that project a magnified image orprofile of a part onto a screen for comparison to a standard overlayprofile or scale; plug/pin gages, which are used for a “go/no-go”assessment of hole and slot dimensions or locations compared tospecified tolerances; protractors and angle gages, which measure theangle between two surfaces of a part or assembly; ring gages, which areused for “go/no-go” assessment compared to the specified dimensionaltolerances or attributes of pins, shafts, or threaded studs; rules andscales, which are flat, graduated scales used for length measurement,and which for OEM applications, digital or electronic linear scales areoften used; snap gages, which are used in production settings wherespecific diametrical or thickness measurements must be repeatedfrequently with precision and accuracy; specialty microscopes, which areused for specialized applications including metallurgy, gemology, or usespecialized techniques like acoustics or microwaves to perform theirfunction; squares, which are used to indicate if two surfaces of a partor assembly are perpendicular; styli, probes, and cantilevers, which areslender rod-shaped stems and contact tips or points used to probesurfaces in conjunction with profilometers, SPMs, CMMs, gages anddimensional scanners; surface profilometers, which measure surfaceprofiles, roughness, waviness and other finish parameters by scanning amechanical stylus across the sample or through noncontact methods;thread gages, which are dimensional instruments for measuring threadsize, pitch or other parameters; and videoscopes, which are inspectiontools that capture images from inside holes, bores or cavities.

XVIII. Open Molding Techniques

According to some embodiments, the particles described herein are formedin an open mold. Open molding can reduce the number of steps andsequences of events required during molding of particles and can improvethe evaporation rate of solvent from the particle precursor material,thereby, increasing the efficiency and rate of particle production.

Referring to FIG. 47, surface or template 4700 includes cavities orrecesses 4702 formed therein. A substance 4704, which can be, but is notlimited to a liquid, a powder, a paste, a gel, a liquified solid,combinations thereof, and the like, is then deposited on surface 4700.The substance 4704 is introduced into recesses 4702 of surface 4700 andexcess substance remaining on surface 4700 is removed 4706. Excesssubstance 4704 can be removed from the surface by, but is not limitedto, doctor blading, applying pressure with a substrate, electrostatics,magnetics, gravitational forces, air pressure, combinations thereof, andthe like. Next, substance 4704 remaining in recesses 4702 is hardenedinto particles 4708 by, but is not limited to, photocuring, thermalcuring, solvent evaporation, oxidation or reductive polymerization,change of temperature, combinations thereof, and the like. Aftersubstance 4704 is hardened, the particles 4708 are harvested fromrecesses 4702.

According to some embodiments, surface 4700 is configured such thatparticle fabrication is accomplished in high throughput. In someembodiments, the surface is configured, for example, planer,cylindrical, spherical, curved, linear, a convery belt type arrangement,a gravure printing type arrangement (such as described in U.S. Pat. Nos.4,557,195 and 4,905,594, all of which are incorporated herein byreference in their entirity), in large sheet arrangements, inmulti-layered sheet arrangements, combinations thereof, and the like.According to such embodiments some recesses in the surface can be in astage of being filled with substance while at another station of thesurface excess substance is being removed. Meanwhile, yet anotherstation of the surface can be hardening the substance and still anotherstation being responsible for harvesting the particles from therecesses. In such embodiments, particles are fabricated efficiently andeffectively in high throughput. In some embodiments the method andsystem are continuous, in other embodiments the method and system arebatch, and in some embodiments the method and system are a combinationof continuous and batch.

The composition of surface 4700 itself can be fabricated from anymaterial that is chemically, physically, and commercially viable for aparticular process to be carried out. According to some embodiments, thematerial for fabrication of surface 4700 is any of the materialsdescribed herein. More particularly, the material of surface 4700 is anymaterial that has a low surface energy, is non-wettable, highlychemically inert, a solvent resistant low surface energy polymericmaterial, a solvent resistant elastomeric material, combinationsthereof, and the like. Even more particularly, the material from whichsurface 4700 is fabricated is a perfluoropolyether material, a siliconematerial, a fluoroolefin material, an acrylate material, a siliconematerial, a styrenic material, a fluorinated thermoplastic elastomer(TPE), a triazine fluoropolymer, a perfluorocyclobutyl material, afluorinated epoxy resin, a fluorinated monomer or fluorinated oligomerthat can be polymerized or crosslinked by a metathesis polymerizationreaction, combinations thereof, and the like.

According to some embodiments, recesses 4702 in surface 4700 arerecesses of particular shapes and sizes. Recesses 4702 can be, but arenot limited to, regular shaped, irregular shaped, variable shaped, andthe like. In some embodiments, recesses 4702 are, but are not limitedto, arched recesses, recesses with right angles, tapered recesses,diamond shaped, spherical, rectangle, triangle, polymorphic, molecularshaped, protein shaped, combinations thereof, and the like. In someembodiments, recesses 4702 can be electrically and/or chemically chargedsuch that functional monomers within substance 4704 are attracted and/orrepelled, thereby resulting in a functional particle as describedelsewhere herein. According to some embodiments, recess 4704 is lessthan about 1 mm in a dimension. According to some embodiments, therecess is less than about 1 mm in its largest cross-sectional dimension.In other embodiments the recess includes a dimension that is betweenabout 20 nm and about 1 mm. In other embodiments, the recess is betweenabout 20 nm and about 500 micron in a dimension and/or in a largestdimension. More particularly, the recess is between about 50 nm andabout 250 micron in a dimension and/or in a largest dimension.

According to embodiments of the present invention, any of the substancesdisclosed herein, for example, a drug, DNA, RNA, a biological molecule,a super absorptive material, combinations thereof, and the like can besubstance 4704 that is deposited into recesses 4702 and molded into aparticle. According to still further embodiments, substance 4704 to bemolded is, but is not limited to, a polymer, a solution, a monomer, aplurality of monomers, a polymerization initiator, a polymerizationcatalyst, an inorganic precursor, a metal precursor, a pharmaceuticalagent, a tag, a magnetic material, a paramagnetic material, a ligand, acell penetrating peptide, a porogen, a surfactant, a plurality ofimmiscible liquids, a solvent, a charged species, combinations thereof,and the like. In still further embodiments, particle 4708 is, but is notlimited to, organic polymers, charged particles, polymer electrets(poly(vinylidene fluoride), Teflon-fluorinated ethylene propylene,polytetrafluoroethylene), therapeutic agents, drugs, non-viral genevectors, RNAi, viral particles, polymorphs, combinations thereof, andthe like.

According to embodiments of the invention, substance 4704 to be moldedinto particles 4708 is deposited onto template surface 4700. In someembodiments substance 4704 is in a liquid form and therefore flows intorecesses 4702 of surface 4700. According to other embodiments, substance4704 takes on another physical form, such as for example, a powder, agel, a paste, or the like, such that a force can be required to ensuresubstance 4704 becomes introduced into recesses 4702. Such a force thatcan be useful in introducing substance 4704 into recesses 4702 can be,but is not limited to, vibration, centrifugal, electrostatic, magnetic,electromagnetic, gravity, compression, combinations thereof, and thelike. The force can also be utilized in embodiments where substance 4704is a liquid to further ensure substance 4704 enters into recesses 4702.

Following introduction of substance 4704 onto template surface 4700 andrecesses 4702 thereof, excess substance is removed from surface 4700 insome embodiments. Removal of excess substance 4704 can be accomplishedby engaging surface 4700 with a second surface 4712 such that the excesssubstance is squeezed out. Second surface 4712 can be, but is notlimited to, a flat surface, an arched surface, and the like. In someembodiments second surface 4712 is brought into contact with templatesurface 4700. According to other embodiments second surface 4712 isbrought within a predetermine distance of template surface 4700.According to some embodiments, second surface 4712 is positioned withrespect to template surface 4700 normal to the plane of template surface4700. According to other embodiments second surface 4712 engagestemplate surface 4700 with a predetermined contact angle. According tostill further embodiments, second surface 4712 can be an arched surface,such as a cylinder, and can be rolled with respect to template surface4700 to remove excess substance. According to yet further embodiments,second surface 4712 is composed of a composition that repells orattracts the excess substance, such as for example, a non-wettingsubstance, a hydrophobic surface repelling a hydrophilic substance, andthe like.

According to other embodiments, excess substance 4704 can be removedfrom template surface 4700 by doctor blading, or otherwise passing ablade across template surface 4700. According to some embodiments, blade4714 is composed of a metal, rubber, polymer, silicon based material,glass, hydrophobic substance, hydrophilic substance, combinationsthereof, and the like. In some embodiments blade 4714 is positioned tocontact surface 4700 and wipe away excess substance. In otherembodiments, blade 4714 is positioned a predetermined distance fromsurface 4700 and drawn across surface 4700 to remove excess substancefrom template surface 4700. The distance blade 4714 is positioned fromsurface 4700 and the rate at which blade 4714 is drawn across surface4700 are variable and determined by the material properties of blade4714, template surface 4700, substance 4704 to be molded, combinationsthereof, and the like. Doctor blading and similar techniques aredisclosed in Lee et al., Two-Polymer Microtransfer Molding for HighlyLayered Microstructures, Adv. Mater. 2005, 17, 2481-2485, which isincorporated herein by reference in its entirity.

Substance 4704 in recesses 4702 is then hardened to form particles 4708.The hardening of substance 4704 can be achieved by any of the methodsand by utilizing any of the materials described herein. According tosome embodiments the hardening is accomplished by, but is not limitedto, solvent evaporation, photo curing, thermal curing, cooling,combinations thereof, and the like.

After substance 4704 has been hardened, particles 4708 are harvestedfrom recesses 4702. According to some embodiments particle 4708 isharvested by contacting particle 4708 with an article that has affinityfor particles 4708 that is greater than the affinity between particle4708 and recess 4702. By way of example, but not limitation, particle4708 is harvested by contacting particle 4708 with an adhesive substancethat adheres to particle 4708 with greater affinity than affinitybetween particle 4708 and template recess 4702. According to someembodiments, the harvesting substance is, but is not limited to, water,organic solvents, carbohydrates, epoxies, waxes, polyvinyl alcohol,polyvinyl pyrrolidone, polybutyl acrylate, polycyano acrylates,polymethyl methacrylate, combinations thereof, and the like. Accordingto still further embodiments substance 4704 in recesses 4702 forms aporous particle by solvent casting.

According to other embodiments, particles 4708 are harvested bysubjecting the particle/recess combination and/or template surface to aphysical force or energy such that particles 4708 are released from therecess 4702. In some embodiments the force is, but is not limited to,centrifugation, dissolution, vibration, ultrasonics, megasonics,gravity, flexure of the template, suction, electrostatic attraction,electrostatic repulsion, magnetism, physical template manipulation,combinations thereof, and the like.

According to some embodiments, particles 4708 are purified after beingharvested. In some embodiments particles 4708 are purified from theharvesting substance. The harvesting can be, but is not limited to,centrifugation, separation, vibration, gravity, dialysis, filtering,sieving, electrophoresis, gas stream, magnetism, electrostaticseparation, combinations thereof, and the like.

XVIII.A. Particles Formed From Open Molding

According to some embodiments, recesses 4702 are sized and shaped suchthat particles formed therefrom will make polymorphs of drugs. Forming adrug from particles 4708 of specific sizes and shapes can increase theefficacy, efficiency, potency, and the like, of a drug substance. Formore on polymorphs, see Lee et al., Crystalliztion on ConfinedEngineered Surfaces: A Method to Control Crystal Size and GenerateDifferent Polymorphs, J. Am. Chem. Soc., 127 (43), 14982-14983, 2005,which is incorporated herein by reference in its entirity.

According to some embodiments, particles 4708 form super absorbentpolymer particles. Examples of super absorbent polymer materials thatcan be made into particles 4708 according to the present invention,include, but are not limited to, polyacrylates, polyacrylic acid,polyacrylamide, cellulose ethers, poly (ethylene oxide), poly (vinylalcohol), polysuccinimides, polyacrylonitrile polymers, combinationsthereof, and the like. According to further embodiments, these superabsorbent polymers can be blended or crosslinked with other polymers, ortheir monomers can be co-polymerized with other monomers, or the like.According to still further embodiments, a starch is grafted onto thesepolymers.

According to further embodiments, particle 4708 formed from the methodsand materials of the present invention include, but are not limited to,particles between 20 nm and 10 microns of a drug, a charged particle, apolymer electret, a therapeutic agent, a viral particle, a polymorph, asuper absorbent particle, combinations thereof, and the like.

XVIV. Seed Coating

According to some embodiments of the present invention, the materialsand methods disclosed herein are used to coat seeds. Referring now toFIG. 48, to coat seeds, the seeds are suspended in a liquid solution4808. The liquid solution containing the seeds 4808 is deposited onto atemplate 4802, where the template includes a recess 4812. The liquidsolution containing the seed 4808 is brought into the recesses 4812 andthe liquid is hardened such that the seed becomes coated. The coatedseeds are then harvested from the recesses 4810. Harvesting of thecoated seeds can be accomplished by any of the harvesting methodsdescribed herein.

According to some embodiments, template 4802 is generated by introducinga liquid template precursor to a scaffolding 4800 which contains apattern that template 4802 will mask. The liquid template precursor isthen hardened to form template 4802. The liquid template precursor canbe any of the materials disclosed herein and can be hardened by any ofthe methods and materials disclosed herein. For example, the liquidtemplate precursor can be a liquid PFPE precursor and contain a curablecomponent (e.g., UV, photo, thermal, combinations thereof, and thelike). According to this example, the liquid PFPE precursor isintroduced to scaffolding 4800 and treated with UV radiation to cure theliquid PFPE into solid form.

According to further embodiments, liquid solution containing the seed4808 is desposited onto a platform 4804 which is configured to sandwichliquid solution 4808 with template 4802. When liquid solution 4808 hasbeen sandwiched into recesses 4812 of template 4802, liquid solutioncontaining the seed 4808 is hardened such that the seed is coated in asolidified material 4810. Hardening can be by any of the methods andsystems described herein, including, but not limited to, photo curing,thermal curing, evaporation, and the like. Following hardening of liquidsolution 4808, platform 4804 and template 4802 are removed from eachother and solidified coated seeds 4810 are harvested from template 4802and/or the surface of platform 4804. Harvesting can be any of theharvesting methods described herein.

The coating of seeds with the materials and methods disclosed hereincan, but is not limited to, preparing the seed for packaging, prepairingcoated seeds of a uniform size, prepairing seeds with a uniform coating,preparing seeds with a uniform coated shape, eliminating surfactants,combinations thereof, and the like. Seed coating techniques compatiblewith the present invention are disclosed in U.S. Pat. No. 4,245,432,which is incorporated herein by reference in its entirity.

EXAMPLES

The following Examples have been included to provide guidance to one ofordinary skill in the art for practicing representative embodiments ofthe presently disclosed subject matter. In light of the presentdisclosure and the general level of skill in the art, those of skill canappreciate that the following Examples are intended to be exemplary onlyand that numerous changes, modifications, and alterations can beemployed without departing from the scope of the presently disclosedsubject matter.

Example 1 Representative Procedure for Synthesis and Curing PhotocurablePerfluoropolyethers

In some embodiments, the synthesis and curing of PFPE materials of thepresently disclosed subject matter is performed by using the methoddescribed by Rolland, J. P., et al., J. Am. Chem. Soc., 2004, 126,2322-2323. Briefly, this method involves themethacrylate-functionalization of a commercially available PFPE diol(M_(n)=3800 g/mol) with isocyanatoethyl methacrylate. Subsequentphotocuring of the material is accomplished through blending with 1 wt %of 2,2-dimethoxy-2-phenylacetophenone and exposure to UV radiation(λ=365 nm).

More particularly, in a typical preparation of perfluoropolyetherdimethacrylate (PFPE DMA), poly(tetrafluoroethyleneoxide-co-difluoromethylene oxide)α,ω diol (ZDOL, average M_(n) ca. 3,800g/mol, 95%, Aldrich Chemical Company, Milwaukee, Wis., United States ofAmerica) (5.7227 g, 1.5 mmol) was added to a dry 50 mL round bottomflask and purged with argon for 15 minutes. 2-isocyanatoethylmethacrylate (EIM, 99%, Aldrich) (0.43 mL, 3.0 mmol) was then added viasyringe along with 1,1,2-trichlorotrifluoroethane (Freon 113 99%,Aldrich) (2 mL), and dibutyltin diacetate (DBTDA, 99%, Aldrich) (50 μL).The solution was immersed in an oil bath and allowed to stir at 50° C.for 24 h. The solution was then passed through a chromatographic column(alumina, Freon 113, 2×5 cm). Evaporation of the solvent yielded aclear, colorless, viscous oil, which was further purified by passagethrough a 0.22-μm polyethersulfone filter.

In a representative curing procedure for PFPE DMA, 1 wt % of2,2-dimethoxy-2-phenyl acetophenone (DMPA, 99% Aldrich), (0.05 g, 2.0mmol) was added to PFPE DMA (5 g, 1.2 mmol) along with 2 mL Freon 113until a clear solution was formed. After removal of the solvent, thecloudy viscous oil was passed through a 0.22-μm polyethersulfone filterto remove any DMPA that did not disperse into the PFPE DMA. The filteredPFPE DMA was then irradiated with a UV source (Electro-Lite Corporation,Danbury, Conn., United States of America, UV curing chamber model no.81432-ELC-500, λ=365 nm) while under a nitrogen purge for 10 min. Thisresulted in a clear, slightly yellow, rubbery material.

Example 2 Representative Fabrication of a PFPE DMA Device

In some embodiments, a PFPE DMA device, such as a stamp, was fabricatedaccording to the method described by Rolland, J. P., et al., J. Am.Chem. Soc., 2004, 126, 2322-2323. Briefly, the PFPE DMA containing aphotoinitiator, such as DMPA, was spin coated (800 rpm) to a thicknessof 20 μm onto a Si wafer containing the desired photoresist pattern.This coated wafer was then placed into the UV curing chamber andirradiated for 6 seconds. Separately, a thick layer (about 5 mm) of thematerial was produced by pouring the PFPE DMA containing photoinitiatorinto a mold surrounding the Si wafer containing the desired photoresistpattern. This wafer was irradiated with UV light for one minute.Following this, the thick layer was removed. The thick layer was thenplaced on top of the thin layer such that the patterns in the two layerswere precisely aligned, and then the entire device was irradiated for 10minutes. Once complete, the entire device was peeled from the Si waferwith both layers adhered together.

Example 3 Fabrication of Isolated Particles using Non-Wetting ImprintLithography

3.1 Fabrication of 200-nm Trapezoidal PEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(See FIG. 13). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus was then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold was then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of PEG diacrylate is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate. The pressure used was atleast about 100 N/cm². The entire apparatus was then subjected to UVlight (λ=365 nm) for ten minutes while under a nitrogen purge. Particlesare observed after separation of the PFPE mold and the treated siliconwafer using scanning electron microscopy (SEM) (see FIG. 14).

3.2 Fabrication of 500-nm Conical PEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of PEG diacrylate is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate. The entire apparatus isthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. Particles are observed after separation of the PFPE moldand the treated silicon wafer using scanning electron microscopy (SEM)(see FIG. 15).

3.3 Fabrication of 3-ρm Arrow-Shaped PEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 3-μm arrow shapes (seeFIG. 11). A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of PEG diacrylate is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate. The entire apparatus isthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. Particles are observed after separation of the PFPE moldand the treated silicon wafer using scanning electron microscopy (SEM)(see FIG. 16).

3.4 Fabrication of 200-nm×750-nm×250-nm Rectangular PEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm×750-nm×250-nmrectangular shapes. A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of PEG diacrylate is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate. The entire apparatus isthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. Particles are observed after separation of the PFPE moldand the treated silicon wafer using scanning electron microscopy (SEM)(see FIG. 17).

3.5 Fabrication of 200-nm Trapezoidal Trimethylopropane Triacrylate(TMPTA) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. Flat, uniform, non-wetting surfacesare generated by treating a silicon wafer cleaned with “piranha”solution (1:1 concentrated sulfuric acid: 30% hydrogen peroxide (aq)solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapordeposition in a desiccator for 20 minutes. Following this, 50 μL ofTMPTA is then placed on the treated silicon wafer and the patterned PFPEmold placed on top of it. The substrate is then placed in a moldingapparatus and a small pressure is applied to push out excess TMPTA. Theentire apparatus is then subjected to UV light (λ=365 nm) for tenminutes while under a nitrogen purge. Particles are observed afterseparation of the PFPE mold and the treated silicon wafer using scanningelectron microscopy (SEM) (see FIG. 18).

3.6 Fabrication of 500-nm Conical Trimethylopropane Triacrylate (TMPTA)Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. Flat, uniform, non-wetting surfacesare generated by treating a silicon wafer cleaned with “piranha”solution (1:1 concentrated sulfuric acid: 30% hydrogen peroxide (aq)solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapordeposition in a desiccator for 20 minutes. Following this, 50 μL ofTMPTA is then placed on the treated silicon wafer and the patterned PFPEmold placed on top of it. The substrate is then placed in a moldingapparatus and a small pressure is applied to push out excess TMPTA. Theentire apparatus is then subjected to UV light (λ=365 nm) for tenminutes while under a nitrogen purge. Particles are observed afterseparation of the PFPE mold and the treated silicon wafer using scanningelectron microscopy (SEM) (see FIG. 19). Further, FIG. 20 shows ascanning electron micrograph of 500-nm isolated conical particles ofTMPTA, which have been printed using an embodiment of the presentlydescribed non-wetting imprint lithography method and harvestedmechanically using a doctor blade. The ability to harvest particles insuch a way offers conclusive evidence for the absence of a “scum layer.”

3.7 Fabrication of 3-μm Arrow-Shaped TMPTA Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 3-μm arrow shapes (seeFIG. 11). A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. Flat, uniform, non-wetting surfacesare generated by treating a silicon wafer cleaned with “piranha”solution (1:1 concentrated sulfuric acid: 30% hydrogen peroxide (aq)solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapordeposition in a desiccator for 20 minutes. Following this, 50 μL ofTMPTA is then placed on the treated silicon wafer and the patterned PFPEmold placed on top of it. The substrate is then placed in a moldingapparatus and a small pressure is applied to push out excess TMPTA. Theentire apparatus is then subjected to UV light (λ=365 nm) for tenminutes while under a nitrogen purge. Particles are observed afterseparation of the PFPE mold and the treated silicon wafer using scanningelectron microscopy (SEM).

3.8 Fabrication of 200-nm Trapezoidal Poly(Lactic Acid) (PLA) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light-(λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, one gram of (3S)-cis-3,6-dimethyl-1,4-dioxane-2,5-dione (LA)is heated above its melting temperature (92° C.) to 110° C. andapproximately 20 μL of stannous octoate catalyst/initiator is added tothe liquid monomer. Flat, uniform, non-wetting surfaces are generated bytreating a silicon wafer cleaned with “piranha” solution (1:1concentrated sulfuric acid: 30% hydrogen peroxide (aq) solution) withtrichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapor deposition ina desiccator for 20 minutes. Following this, 50 μL of molten LAcontaining catalyst is then placed on the treated silicon waferpreheated to 110° C. and the patterned PFPE mold is placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess monomer. The entire apparatus is thenplaced in an oven at 110° C. for 15 hours. Particles are observed aftercooling to room temperature and separation of the PFPE mold and thetreated silicon wafer using scanning electron microscopy (SEM) (see FIG.21). Further, FIG. 22 is a scanning electron micrograph of 200-nmisolated trapezoidal particles of poly(lactic acid) (PLA), which havebeen printed using an embodiment of the presently described non-wettingimprint lithography method and harvested mechanically using a doctorblade. The ability to harvest particles in such a way offers conclusiveevidence for the absence of a “scum layer.”

3.9 Fabrication of 3-μm Arrow-Shaped (PLA) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 3-μm arrow shapes (seeFIG. 11). A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, one gram of (3S)-cis-3,6-dimethyl-1,4-dioxane-2,5-dione (LA)is heated above its melting temperature (92° C.) to 110° C. andapproximately 20 μL of stannous octoate catalyst/initiator is added tothe liquid monomer. Flat, uniform, non-wetting surfaces are generated bytreating a silicon wafer cleaned with “piranha” solution (1:1concentrated sulfuric acid: 30% hydrogen peroxide (aq) solution) withtrichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapor deposition ina desiccator for 20 minutes. Following this, 50 μL of molten LAcontaining catalyst is then placed on the treated silicon waferpreheated to 110° C. and the patterned PFPE mold is placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess monomer. The entire apparatus is thenplaced in an oven at 110° C. for 15 hours. Particles are observed aftercooling to room temperature and separation of the PFPE mold and thetreated silicon wafer using scanning electron microscopy (SEM) (see FIG.23).

3.10 Fabrication of 500-nm Conical Shaped (PLA) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, one gram of (3S)-cis-3,6-dimethyl-1,4-dioxane-2,5-dione (LA)is heated above its melting temperature (92° C.) to 110° C. andapproximately 20 μL of stannous octoate catalyst/initiator is added tothe liquid monomer. Flat, uniform, non-wetting surfaces are generated bytreating a silicon wafer cleaned with “piranha” solution (1:1concentrated sulfuric acid: 30% hydrogen peroxide (aq) solution) withtrichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapor deposition ina desiccator for 20 minutes. Following this, 50 μL of molten LAcontaining catalyst is then placed on the treated silicon waferpreheated to 110° C. and the patterned PFPE mold is placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess monomer. The entire apparatus is thenplaced in an oven at 110° C. for 15 hours. Particles are observed aftercooling to room temperature and separation of the PFPE mold and thetreated silicon wafer using scanning electron microscopy (SEM) (see FIG.24).

3.11 Fabrication of 200-nm Trapezoidal Poly(pyrrole) (Ppy) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, 50 μL of a 1:1 v:v solution oftetrahydrofuran:pyrrole is added to 50 μL of 70% perchloric acid (aq). Aclear, homogenous, brown solution quickly forms and develops into black,solid, polypyrrole in 15 minutes. A drop of this clear, brown solution(prior to complete polymerization) is placed onto a treated siliconwafer and into a stamping apparatus and a pressure is applied to removeexcess solution. The apparatus is then placed into a vacuum oven for 15h to remove the THF and water. Particles are observed using scanningelectron microscopy (SEM) (see FIG. 25) after release of the vacuum andseparation of the PFPE mold and the treated silicon wafer.

3.12 Fabrication of 3-μm Arrow-Shaped (Ppy) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 3-μm arrow shapes (seeFIG. 11). A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master. Flat,uniform, non-wetting surfaces are generated by treating a silicon wafercleaned with “piranha” solution (1:1 concentrated sulfuric acid: 30%hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, 50 μL of a 1:1 v:v solution oftetrahydrofuran:pyrrole is added to 50 μL of 70% perchloric acid (aq). Aclear, homogenous, brown solution quickly forms and develops into black,solid, polypyrrole in 15 minutes. A drop of this clear, brown solution(prior to complete polymerization) is placed onto a treated siliconwafer and into a stamping apparatus and a pressure is applied to removeexcess solution. The apparatus is then placed into a vacuum oven for 15h to remove the THF and water. Particles are observed using scanningelectron microscopy (SEM) (see FIG. 26) after release of the vacuum andseparation of the PFPE mold and the treated silicon wafer.

3.13 Fabrication of 500-nm Conical (Ppy) Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, 50 μL of a 1:1 v:v solution oftetrahydrofuran:pyrrole is added to 50 μL of 70% perchloric acid (aq). Aclear, homogenous, brown solution quickly forms and develops into black,solid, polypyrrole in 15 minutes. A drop of this clear, brown solution(prior to complete polymerization) is placed onto a treated siliconwafer and into a stamping apparatus and a pressure is applied to removeexcess solution. The apparatus is then placed into a vacuum oven for 15h to remove the THF and water. Particles are observed using scanningelectron microscopy (SEM) (see FIG. 27) after release of the vacuum andseparation of the PFPE mold and the treated silicon wafer.

3.14 Encapsulation of Fluorescently Tagged DNA Inside 200-nm TrapezoidalPEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone. 20μL of water and 20 μL of PEG diacrylate monomer are added to 8 nanomolesof 24 bp DNA oligonucleotide that has been tagged with a fluorescentdye, CY-3. Flat, uniform, non-wetting surfaces are generated by treatinga silicon wafer cleaned with “piranha” solution (1:1 concentratedsulfuric acid: 30% hydrogen peroxide (aq) solution) with trichloro(1H,1H, 2H, 2H-perfluorooctyl) silane via vapor deposition in a desiccatorfor 20 minutes. Following this, 50 μL of the PEG diacrylate solution isthen placed on the treated silicon wafer and the patterned PFPE moldplaced on top of it. The substrate is then placed in a molding apparatusand a small pressure is applied to push out excess PEG-diacrylatesolution. The entire apparatus is then subjected to UV light (λ=365 nm)for ten minutes while under a nitrogen purge. Particles are observedafter separation of the PFPE mold and the treated silicon wafer usingconfocal fluorescence microscopy (see FIG. 28). Further, FIG. 28A showsa fluorescent confocal micrograph of 200-nm trapezoidal PEGnanoparticles which contain 24-mer DNA strands that are tagged withCY-3. FIG. 28B is optical micrograph of the 200-nm isolated trapezoidalparticles of PEG diacrylate that contain fluorescently tagged DNA. FIG.28C is the overlay of the images provided in FIGS. 28A and 28B, showingthat every particle contains DNA.

3.15 Encapsulation of Magnetite Nanoparticles Inside 500-nm Conical PEGParticles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, citrate capped magnetite nanoparticles weresynthesized by reaction of ferric chloride (40 mL of a 1 M aqueoussolution) and ferrous chloride (10 mL of a 2 M aqueous hydrochloric acidsolution) which is added to ammonia (500 mL of a 0.7 M aqueoussolution). The resulting precipitate is collected by centrifugation andthen stirred in 2 M perchloric acid. The final solids are collected bycentrifugation. 0.290 g of these perchlorate-stabilized nanoparticlesare suspended in 50 mL of water and heated to 90° C. while stirring.Next, 0.106 g of sodium citrate is added. The solution is stirred at 90°C. for 30 min to yield an aqueous solution of citrate-stabilized ironoxide nanoparticles. 50 μL of this solution is added to 50 μL of a PEGdiacrylate solution in a microtube. This microtube is vortexed for tenseconds. Following this, 50 μL of this PEG diacrylate/particle solutionis then placed on the treated silicon wafer and the patterned PFPE moldplaced on top of it. The substrate is then placed in a molding apparatusand a small pressure is applied to push out excessPEG-diacrylate/particle solution. The entire apparatus is then subjectedto UV light (λ=365 nm) for ten minutes while under a nitrogen purge.Nanoparticle-containing PEG-diacrylate particles are observed afterseparation of the PFPE mold and the treated silicon wafer using opticalmicroscopy.

3.16 Fabrication of Isolated Particles on Glass Surfaces Using “DoubleStamping”

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone. Aflat, non-wetting surface is generated by photocuring a film of PFPE-DMAonto a glass slide, according to the procedure outlined for generating apatterned PFPE-DMA mold. 5 μL of the PEG-diacrylate/photoinitiatorsolution is pressed between the PFPE-DMA mold and the flat PFPE-DMAsurface, and pressure is applied to squeeze out excess PEG-diacrylatemonomer. The PFPE-DMA mold is then removed from the flat PFPE-DMAsurface and pressed against a clean glass microscope slide andphotocured using UV radiation (λ=365 nm) for 10 minutes while under anitrogen purge. Particles are observed after cooling to room temperatureand separation of the PFPE mold and the glass microscope slide, usingscanning electron microscopy (SEM) (see FIG. 29).

3.17 Encapsulation of Viruses in PEG-Diacrylate Nanoparticles

A patterned perfluoropolyether (PFPE) mold is generated by pouringPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Fluorescently-labeled or unlabeled Adenovirus or Adeno-Associated Virussuspensions are added to this PEG-diacrylate monomer solution and mixedthoroughly. Flat, uniform, non-wetting surfaces are generated bytreating a silicon wafer cleaned with “piranha” solution (1:1concentrated sulfuric acid: 30% hydrogen peroxide (aq) solution) withtrichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapor deposition ina desiccator for 20 minutes. Following this, 50 μL of the PEGdiacrylate/virus solution is then placed on the treated silicon waferand the patterned PFPE mold placed on top of it. The substrate is thenplaced in a molding apparatus and a small pressure is applied to pushout excess PEG-diacrylate solution. The entire apparatus is thensubjected to UV light (λ=365 nm) for ten minutes while under a nitrogenpurge. Virus-containing particles are observed after separation of thePFPE mold and the treated silicon wafer using transmission electronmicroscopy or, in the case of fluorescently-labeled viruses, confocalfluorescence microscopy.

3.18 Encapsulation of Proteins in PEG-Diacrylate Nanoparticles

A patterned perfluoropolyether (PFPE) mold is generated by pouringPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Fluorescently-labeled or unlabeled protein solutions are added to thisPEG-diacrylate monomer solution and mixed thoroughly. Flat, uniform,non-wetting surfaces are generated by treating a silicon wafer cleanedwith “piranha” solution (1:1 concentrated sulfuric acid: 30% hydrogenperoxide (aq) solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl)silane via vapor deposition in a desiccator for 20 minutes. Followingthis, 50 μL of the PEG diacrylate/virus solution is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate solution. The entireapparatus is then subjected to UV light (λ=365 nm) for ten minutes whileunder a nitrogen purge. Protein-containing particles are observed afterseparation of the PFPE mold and the treated silicon wafer usingtraditional assay methods or, in the case of fluorescently-labeledproteins, confocal fluorescence microscopy.

3.19 Fabrication of 200-nm Titania Particles

A patterned perfluoropolyether (PFPE) mold can be generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidalshapes, such as shown in FIG. 13. A poly(dimethylsiloxane) mold can beused to confine the liquid PFPE-DMA to the desired area. The apparatuscan then be subjected to UV light (λ=365 nm) for 10 minutes while undera nitrogen purge. The fully cured PFPE-DMA mold is then released fromthe silicon master. Separately, 1 g of Pluronic P123 is dissolved in 12g of absolute ethanol. This solution was added to a solution of 2.7 mLof concentrated hydrochloric acid and 3.88 mL titanium (IV) ethoxide.Flat, uniform, non-wetting surfaces can be generated by treating asilicon wafer cleaned with “piranha” solution (1:1 concentrated sulfuricacid: 30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of the sol-gel solution can then beplaced on the treated silicon wafer and the patterned PFPE mold placedon top of it. The substrate is then placed in a molding apparatus and asmall pressure is applied to push out excess sol-gel precursor. Theentire apparatus is then set aside until the sol-gel precursor hassolidified. After solidification of the sol-gel precursor, the siliconwafer can be removed from the patterned PFPE and particles will bepresent.

3.20 Fabrication of 200-nm Silica Particles

A patterned perfluoropolyether (PFPE) mold can be generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidalshapes, such as shown in FIG. 13. A poly(dimethylsiloxane) mold can thenbe used to confine the liquid PFPE-DMA to the desired area. Theapparatus can then be subjected to UV light (λ=365 nm) for 10 minuteswhile under a nitrogen purge. The fully cured PFPE-DMA mold is thenreleased from the silicon master. Separately, 2 g of Pluronic P123 isdissolved in 30 g of water and 120 g of 2 M HCl is added while stirringat 35° C. To this solution, add 8.50 g of TEOS with stirring at 35° C.for 20 h. Flat, uniform, non-wetting surfaces can then be generated bytreating a silicon wafer cleaned with “piranha” solution (1:1concentrated sulfuric acid: 30% hydrogen peroxide (aq) solution) withtrichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapor deposition ina desiccator for 20 minutes. Following this, 50 μL of the sol-gelsolution is then placed on the treated silicon wafer and the patternedPFPE mold placed on top of it. The substrate is then placed in a moldingapparatus and a small pressure is applied to push out excess sol-gelprecursor. The entire apparatus is then set aside until the sol-gelprecursor has solidified. Particles should be observed after separationof the PFPE mold and the treated silicon wafer using scanning electronmicroscopy (SEM).

3.21 Fabrication of 200-nm Europium-Doped Titania Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, 1 g of Pluronic P123 and 0.51 g of EuCl₃.6H₂O are dissolvedin 12 g of absolute ethanol. This solution is added to a solution of 2.7mL of concentrated hydrochloric acid and 3.88 mL titanium (IV) ethoxide.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of the sol-gel solution is then placed onthe treated silicon wafer and the patterned PFPE mold placed on top ofit. The substrate is then placed in a molding apparatus and a smallpressure is applied to push out excess sol-gel precursor. The entireapparatus is then set aside until the sol-gel precursor has solidified.Next, after the sol-gel precursor has solidified, the PFPE mold and thetreated silicon wafer are separated and particles should be observedusing scanning electron microscopy (SEM).

3.22 Encapsulation of CdSe Nanoparticles Inside 200-nm PEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, 0.5 g of sodium citrate and 2 mL of 0.04 M cadmiumperchlorate are dissolved in 45 mL of water, and the pH is adjusted toof the solution to 9 with 0.1 M NaOH. The solution is bubbled withnitrogen for 15 minutes. 2 mL of 1 M N,N-dimethylselenourea is added tothe solution and heated in a microwave oven for 60 seconds. 50 μL ofthis solution is added to 50 μL of a PEG diacrylate solution in amicrotube. This microtube is vortexed for ten seconds. 50 μL of this PEGdiacrylate/CdSe particle solution is placed on the treated silicon waferand the patterned PFPE mold placed on top of it. The substrate is thenplaced in a molding apparatus and a small pressure is applied to pushout excess PEG-diacrylate solution. The entire apparatus is thensubjected to UV light (λ=365 nm) for ten minutes while under a nitrogenpurge. PEG-diacrylate particles with encapsulated CdSe nanoparticleswill be observed after separation of the PFPE mold and the treatedsilicon wafer using TEM or fluorescence microscopy.

3.23 Synthetic Replication of Adenovirus Particles Using Non-WettingImprint Lithography

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing adenovirusparticles on a silicon wafer. This master can be used to template apatterned mold by pouring PFPE-DMA containing 1-hydroxycyclohexyl phenylketone over the patterned area of the master. A poly(dimethylsiloxane)mold is used to confine the liquid PFPE-DMA to the desired area. Theapparatus is then subjected to UV light (λ=365 nm) for 10 minutes whileunder a nitrogen purge. The fully cured PFPE-DMA mold is then releasedfrom the master. Separately, TMPTA is blended with 1 wt % of aphotoinitiator, 1-hydroxycyclohexyl phenyl ketone. Flat, uniform,non-wetting surfaces are generated by treating a silicon wafer cleanedwith “piranha” solution (1:1 concentrated sulfuric acid: 30% hydrogenperoxide (aq) solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl)silane via vapor deposition in a desiccator for 20 minutes. Followingthis, 50 μL of TMPTA is then placed on the treated silicon wafer and thepatterned PFPE mold placed on top of it. The substrate is then placed ina molding apparatus and a small pressure is applied to push out excessTMPTA. The entire apparatus is then subjected to UV light (λ=365 nm) forten minutes while under a nitrogen purge. Synthetic virus replicates areobserved after separation of the PFPE mold and the treated silicon waferusing scanning electron microscopy (SEM) or transmission electronmicroscopy (TEM).

3.24 Synthetic Replication of Earthworm Hemoglobin Protein UsingNon-Wetting Imprint Lithography

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing earthwormhemoglobin protein on a silicon wafer. This master can be used totemplate a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. Separately, TMPTA is blended with1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone. Flat,uniform, non-wetting surfaces are generated by treating a silicon wafercleaned with “piranha” solution (1:1 concentrated sulfuric acid: 30%hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of TMPTA is then placed on the treatedsilicon wafer and the patterned PFPE mold placed on top of it. Thesubstrate is then placed in a molding apparatus and a small pressure isapplied to push out excess TMPTA. The entire apparatus is then subjectedto UV light (λ=365 nm) for ten minutes while under a nitrogen purge.Synthetic protein replicates are observed after separation of the PFPEmold and the treated silicon wafer using scanning electron microscopy(SEM) or transmission electron microscopy (TEM).

3.25 Combinatorial Engineering of 100-nm Nanoparticle Therapeutics

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 100-nm cubic shapes. Apoly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the silicon master. Separately, apoly(ethylene glycol) (PEG) diacrylate (n=9) is blended with 1 wt % of aphotoinitiator, 1-hydroxycyclohexyl phenyl ketone. Other therapeuticagents (i.e., small molecule drugs, proteins, polysaccharides, DNA,etc.), tissue targeting agents (cell penetrating peptides and ligands,hormones, antibodies, etc.), therapeutic release/transfection agents(other controlled-release monomer formulations, cationic lipids, etc.),and miscibility enhancing agents (cosolvents, charged monomers, etc.)are added to the polymer precursor solution in a combinatorial manner.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuricacid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of the combinatorially-generated particleprecursor solution is then placed on the treated silicon wafer and thepatterned PFPE mold placed on top of it. The substrate is then placed ina molding apparatus and a small pressure is applied to push out excesssolution. The entire apparatus is then subjected to UV light (λ=365 nm)for ten minutes while under a nitrogen purge. The PFPE-DMA mold is thenseparated from the treated wafer, particles can be harvested, and thetherapeutic efficacy of each combinatorially generated nanoparticle isestablished. By repeating this methodology with different particleformulations, many combinations of therapeutic agents, tissue targetingagents, release agents, and other important compounds can be rapidlyscreened to determine the optimal combination for a desired therapeuticapplication.

3.26 Fabrication of a Shape-Specific PEG Membrane

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 3-μm cylindrical holesthat are 5 μm deep. A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuricacid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Following this, 50 μL of PEG diacrylate is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to push out excess PEG-diacrylate. The entire apparatus isthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. An interconnected membrane will be observed afterseparation of the PFPE mold and the treated silicon wafer using scanningelectron microscopy (SEM). The membrane will release from the surface bysoaking in water and allowing it to lift off the surface.

3.27 Harvesting of PEG Particles by Ice Formation

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 5-μm cylinder shapes. Thesubstrate is then subjected to a nitrogen purge for 10 minutes, then UVlight (λ=365 nm) is applied for 10 minutes while under a nitrogen purge.The fully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by coating a glassslide with PFPE-DMA containing 1-hydroxycyclohexyl phenyl ketone. Theslide is then subjected to a nitrogen purge for 10 minutes, then UVlight (λ=365 nm) is applied for 10 minutes while under a nitrogen purge.The flat, fully cured PFPE-DMA substrate is released from the slide.Following this, 0.1 mL of PEG diacrylate is then placed on the flatPFPE-DMA substrate and the patterned PFPE mold placed on top of it. Thesubstrate is then placed in a molding apparatus and a small pressure isapplied to push out excess PEG-diacrylate. The entire apparatus is thenpurged with nitrogen for 10 minutes, then subjected to UV light (λ=365nm) for 10 minutes while under a nitrogen purge. PEG particles areobserved after separation of the PFPE-DMA mold and substrate usingoptical microscopy. Water is applied to the surface of the substrate andmold containing particles. A gasket is used to confine the water to thedesired location. The apparatus is then placed in the freezer at atemperature of −10° C. for 30 minutes. The ice containing PEG particlesis peeled off the PFPE-DMA mold and substrate and allowed to melt,yielding an aqueous solution containing PEG particles.

3.28 Harvesting of PEG Particles with Vinyl Pyrrolidone

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 5-μm cylinder shapes. Thesubstrate is then subjected to a nitrogen purge for 10 minutes, then UVlight (λ=365 nm) is applied for 10 minutes while under a nitrogen purge.The fully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by coating a glassslide with PFPE-DMA containing 1-hydroxycyclohexyl phenyl ketone. Theslide is then subjected to a nitrogen purge for 10 minutes, then UVlight (λ=365 nm) is applied for 10 minutes while under a nitrogen purge.The flat, fully cured PFPE-DMA substrate is released from the slide.Following this, 0.1 mL of PEG diacrylate is then placed on the flatPFPE-DMA substrate and the patterned PFPE mold placed on top of it. Thesubstrate is then placed in a molding apparatus and a small pressure isapplied to push out excess PEG-diacrylate. The entire apparatus is thenpurged with nitrogen for 10 minutes, then subjected to UV light (λ=365nm) for 10 minutes while under a nitrogen purge. PEG particles areobserved after separation of the PFPE-DMA mold and substrate usingoptical microscopy. In some embodiments, the material includes anadhesive or sticky surface. In some embodiments, the material includescarbohydrates, epoxies, waxes, polyvinyl alcohol, polyvinyl pyrrolidone,polybutyl acrylate, polycyano acrylates, polymethyl methacrylate. Insome embodiments, the harvesting or collecting of the particles includescooling water to form ice (e.g., in contact with the particles) drop ofn-vinyl-2-pyrrolidone containing 5% photoinitiator, 1-hydroxycyclohexylphenyl ketone, is placed on a clean glass slide. The PFPE-DMA moldcontaining particles is placed patterned side down on then-vinyl-2-pyrrolidone drop. The slide is subjected to a nitrogen purgefor 5 minutes, then UV light (λ=365 nm) is applied for 5 minutes whileunder a nitrogen purge. The slide is removed, and the mold is peeledaway from the polyvinyl pyrrolidone and particles. Particles on thepolyvinyl pyrrolidone were observed with optical microscopy. Thepolyvinyl pyrrolidone film containing particles was dissolved in water.Dialysis was used to remove the polyvinyl pyrrolidone, leaving anaqueous solution containing 5 μm PEG particles.

3.29 Harvesting of PEG Particles with Polyvinyl Alcohol

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 5-μm cylinder shapes. Thesubstrate is then subjected to a nitrogen purge for 10 minutes, then UVlight (λ=365 nm) is applied for 10 minutes while under a nitrogen purge.The fully cured PFPE-DMA mold is then released from the silicon master.Separately, a poly(ethylene glycol) (PEG) diacrylate (n=9) is blendedwith 1 wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone.Flat, uniform, non-wetting surfaces are generated by coating a glassslide with PFPE-DMA containing 1-hydroxycyclohexyl phenyl ketone. Theslide is then subjected to a nitrogen purge for 10 minutes, then UVlight (λ=365 nm) is applied for 10 minutes while under a nitrogen purge.The flat, fully cured PFPE-DMA substrate is released from the slide.Following this, 0.1 mL of PEG diacrylate is then placed on the flatPFPE-DMA substrate and the patterned PFPE mold placed on top of it. Thesubstrate is then placed in a molding apparatus and a small pressure isapplied to push out excess PEG-diacrylate. The entire apparatus is thenpurged with nitrogen for 10 minutes, then subjected to UV light (λ=365nm) for 10 minutes while under a nitrogen purge. PEG particles areobserved after separation of the PFPE-DMA mold and substrate usingoptical microscopy. Separately, a solution of 5 weight percent polyvinylalcohol (PVOH) in ethanol (EtOH) is prepared. The solution is spincoated on a glass slide and allowed to dry. The PFPE-DMA mold containingparticles is placed patterned side down on the glass slide and pressureis applied. The mold is then peeled away from the PVOH and particles.Particles on the PVOH were observed with optical microscopy. The PVOHfilm containing particles was dissolved in water. Dialysis was used toremove the PVOH, leaving an aqueous solution containing 5 μm PEGparticles.

3.30 Fabrication of 200 nm Phosphatidylcholine Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200-nm trapezoidal shapes(see FIG. 13). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toa nitrogen purge for 10 minutes followed by UV light (λ=365 nm) for 10minutes while under a nitrogen purge. The fully cured PFPE-DMA mold isthen released from the silicon master. Separately, flat, uniform,non-wetting surfaces are generated by treating a silicon wafer cleanedwith “piranha” solution (1:1 concentrated sulfuric acid: 30% hydrogenperoxide (aq) solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl)silane via vapor deposition in a desiccator for 20 minutes. Followingthis, 20 mg of the phosphatidylcholine was placed on the treated siliconwafer and heated to 60 degrees C. The substrate is then placed in amolding apparatus and a small pressure is applied to push out excessphosphatidylcholine. The entire apparatus is then set aside until thephosphatidylcholine has solidified. Particles are observed afterseparation of the PFPE mold and the treated silicon wafer using scanningelectron microscopy (SEM).

3.31 Functionalizing PEG Particles with FITC

Poly(ethylene glycol) (PEG) particles with 5 weight percent aminoethylmethacrylate were created. Particles are observed in the PFPE mold afterseparation of the PFPE mold and the PFPE substrate using opticalmicroscopy. Separately, a solution containing 10 weight percentfluorescein isothiocyanate (FITC) in dimethylsulfoxide (DMSO) wascreated. Following this, the mold containing the particles was exposedto the FITC solution for one hour. Excess FITC was rinsed off the moldsurface with DMSO followed by deionized (DI) water. The tagged particleswere observed with fluorescence microscopy, with an excitationwavelength of 492 nm and an emission wavelength of 529 nm.

3.32 Encapsulation of Doxorubicin Inside 500 nm Conical PEG Particles

A patterned perfluoropolyether (PFPE) mold was generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12). A poly(dimethylsiloxane) mold was used to confine theliquid PFPE-DMA to the desired area. The apparatus was then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold was then released from the silicon master.Flat, uniform, non-wetting surfaces were generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, a solution of 1 wt % doxorubicin in PEG diacrylatewas formulated with 1 wt % photoinitiator. Following this, 50 μL of thisPEG diacrylate/doxorubicin solution was then placed on the treatedsilicon wafer and the patterned PFPE mold placed on top of it. Thesubstrate was then placed in a molding apparatus and a small pressurewas applied to push out excess PEG-diacrylate/doxorubicin solution. Thesmall pressure in this example was at least about 100 N/cm². The entireapparatus was then subjected to UV light (λ=365 nm) for ten minuteswhile under a nitrogen purge. Doxorubicin-containing PEG-diacrylateparticles were observed after separation of the PFPE mold and thetreated silicon wafer using fluorescent microscopy (FIG. 42).

3.33 Encapsulation of Avidin (66 kDa) in 160 nm PEG Particles

A patterned perfluoropolyether (PFPE) mold was generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 160-nm cylindrical shapes(see FIG. 43). A poly(dimethylsiloxane) mold was used to confine theliquid PFPE-DMA to the desired area. The apparatus was then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold was then released from the silicon master.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, a solution of 1 wt % avidin in 30:70 PEGmonomethacrylate:PEG diacrylate was formulated with 1 wt %photoinitiator. Following this, 50 μL of this PEG/avidin solution wasthen placed on the treated silicon wafer and the patterned PFPE moldplaced on top of it. The substrate was then placed in a moldingapparatus and a small pressure is applied to push out excessPEG-diacrylate/avidin solution. The small pressure in this example wasat least about 100 N/cm². The entire apparatus was then subjected to UVlight (λ=365 nm) for ten minutes while under a nitrogen purge.Avidin-containing PEG particles were observed after separation of thePFPE mold and the treated silicon wafer using fluorescent microscopy.

3.34 Encapsulation of 2-fluoro-2-deoxy-d-glucose in 80 nm PEG Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a 6 inch silicon substrate patterned with 80-nm cylindricalshapes. The substrate is then subjected to UV light (λ=365 nm) for 10minutes while under a nitrogen purge. The fully cured PFPE-DMA mold isthen released from the silicon master. Flat, uniform, non-wettingsurfaces are generated by treating a silicon wafer cleaned with“piranha” solution (1:1 concentrated sulfuric acid: 30% hydrogenperoxide (aq) solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl)silane via vapor deposition in a desiccator for 20 minutes. Separately,a solution of 0.5 wt % 2-fluoro-2-deoxy-d-glucose (FDG) in 30:70 PEGmonomethacrylate:PEG diacrylate is formulated with 1 wt %photoinitiator. Following this, 200 μL of this PEG/FDG solution is thenplaced on the treated silicon wafer and the patterned PFPE mold placedon top of it. The substrate is then placed in a molding apparatus and asmall pressure is applied to push out excess PEG/FDG solution. The smallpressure should be at least about 100 N/cm². The entire apparatus isthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. FDG-containing PEG particles will be observed afterseparation of the PFPE mold and the treated silicon wafer using scanningelectron microscopy.

3.35 Encapsulated DNA in 200 nm×200 nm×1 μm Bar-Shaped Poly(lactic Acid)Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 200 nm×200 nm×1 μm barshapes. The substrate is then subjected to UV light (λ=365 nm) for 10minutes while under a nitrogen purge. The fully cured PFPE-DMA mold isthen released from the silicon master. Flat, uniform, non-wettingsurfaces are generated by treating a silicon wafer cleaned with“piranha” solution (1:1 concentrated sulfuric acid: 30% hydrogenperoxide (aq) solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl)silane via vapor deposition in a desiccator for 20 minutes. Separately,a solution of 0.01 wt % 24 base pair DNA and 5 wt % poly(lactic acid) inethanol is formulated. 200 μL of this ethanol solution is then placed onthe treated silicon wafer and the patterned PFPE mold placed on top ofit. The substrate is then placed in a molding apparatus and a smallpressure is applied to push out excess PEG/FDG solution. The smallpressure should be at least about 100 N/cm². The entire apparatus isthen placed under vacuum for 2 hours. DNA-containing poly(lactic acid)particles will be observed after separation of the PFPE mold and thetreated silicon wafer using optical microscopy.

3.36 100 nm Paclitaxel Particles

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 500-nm conical shapes(see FIG. 12). A poly(dimethylsiloxane) mold is used to confine theliquid PFPE-DMA to the desired area. The apparatus is then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold is then released from the silicon master.Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, a solution of 5 wt % paclitaxel in ethanol wasformulated. Following this, 100 μL of this paclitaxel solution is thenplaced on the treated silicon wafer and the patterned PFPE mold placedon top of it. The substrate is then placed in a molding apparatus and asmall pressure is applied to push out excess solution. The pressureapplied was at least about 100 N/cm². The entire apparatus is thenplaced under vacuum for 2 hours. Separation of the mold and surfaceyielded approximately 100 nm spherical paclitaxel particles, which wereobserved with scanning electron microscopy.

3.37 Triangular Particles Functionalized on One Side

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a 6 inch silicon substrate patterned with 0.6 μm×0.8 μm×1 μmright triangles. The substrate is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the silicon master. Flat, uniform,non-wetting surfaces are generated by treating a silicon wafer cleanedwith “piranha” solution (1:1 concentrated sulfuric acid: 30% hydrogenperoxide (aq) solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl)silane via vapor deposition in a desiccator for 20 minutes. Separately,a solution of 5 wt % aminoethyl methacrylate in 30:70 PEGmonomethacrylate:PEG diacrylate is formulated with 1 wt %photoinitiator. Following this, 200 μL of this monomer solution is thenplaced on the treated silicon wafer and the patterned PFPE mold placedon top of it. The substrate is then placed in a molding apparatus and asmall pressure is applied to push out excess solution. The smallpressure should be at least about 100 N/cm². The entire apparatus isthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. Aminoethyl methacrylate-containing PEG particles areobserved in the mold after separation of the PFPE mold and the treatedsilicon wafer using optical microscopy. Separately, a solutioncontaining 10 weight percent fluorescein isothiocyanate (FITC) indimethylsulfoxide (DMSO) is created. Following this, the mold containingthe particles is exposed to the FITC solution for one hour. Excess FITCis rinsed off the mold surface with DMSO followed by deionized (DI)water. Particles, tagged only on one face, will be observed withfluorescence microscopy, with an excitation wavelength of 492 nm and anemission wavelength of 529 nm.

3.38 Formation of an Imprinted Protein Binding Cavity and an ArtificialProtein

The desired protein molecules are adsorbed onto a mica substrate tocreate a master template. A mixture of PFPE-dimethacrylate (PFPE-DMA)containing a monomer with a covalently attached disaccharide, and1-hydroxycyclohexyl phenyl ketone as a photoinitiator was poured overthe substrate. The substrate is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the mica master, creating polysaccharide-likecavities that exhibit selective recognition for the protein moleculethat was imprinted. The polymeric mold was soaked in NaOH/NaClO solutionto remove the template proteins.

Flat, uniform, non-wetting surfaces are generated by treating a siliconwafer cleaned with “piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) with trichloro(1H, 1H, 2H,2H-perfluorooctyl) silane via vapor deposition in a desiccator for 20minutes. Separately, a solution of 25% (w/w) methacrylic acid (MAA), 25%diethyl aminoethylmethacrylate (DEAEM), and 48% PEG diacrylate wasformulated with 2 wt % photoinitiator. Following this, 200 μL of thismonomer solution is then placed on the treated silicon wafer and thepatterned PFPE/disaccharide mold placed on top of it. The substrate isthen placed in a molding apparatus and a small pressure is applied topush out excess solution. The entire apparatus is then subjected to UVlight (λ=365 nm) for ten minutes while under a nitrogen purge. Removalof the mold yields artificial protein molecules which have similar size,shape, and chemical functionality as the original template proteinmolecule.

Example 4 Molding of Features for Semiconductor Applications

4.1 Fabrication of 140-nm Lines Separated by 70 nm in TMPTA

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. Flat, uniform, surfaces are generatedby treating a silicon wafer cleaned with “piranha” solution (1:1concentrated sulfuric acid:30% hydrogen peroxide (aq) solution) andtreating the wafer with an adhesion promoter, (trimethoxysilyl propylmethacryalte). Following this, 50 μL of TMPTA is then placed on thetreated silicon wafer and the patterned PFPE mold placed on top of it.The substrate is then placed in a molding apparatus and a small pressureis applied to ensure a conformal contact. The entire apparatus is thensubjected to UV light (λ=365 nm) for ten minutes while under a nitrogenpurge. Features are observed after separation of the PFPE mold and thetreated silicon wafer using atomic force microscopy (AFM) (see FIG. 30).

4.2 Molding of a Polystyrene Solution

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, polystyrene is dissolved in 1 to 99 wt % of toluene. Flat,uniform, surfaces are generated by treating a silicon wafer cleaned with“piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide(aq) solution) and treating the wafer with an adhesion promoter.Following this, 50 μL of polystyrene solution is then placed on thetreated silicon wafer and the patterned PFPE mold is placed on top ofit. The substrate is then placed in a molding apparatus and a smallpressure is applied to ensure a conformal contact. The entire apparatusis then subjected to vacuum for a period of time to remove the solvent.Features are observed after separation of the PFPE mold and the treatedsilicon wafer using atomic force microscopy (AFM) and scanning electronmicroscopy (SEM).

4.3 Molding of Isolated Features on Microelectronics-Compatible Surfacesusing “double Stamping”

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. A flat, non-wetting surface isgenerated by photocuring a film of PFPE-DMA onto a glass slide,according to the procedure outlined for generating a patterned PFPE-DMAmold. 50 μL of the TMPTA/photoinitiator solution is pressed between thePFPE-DMA mold and the flat PFPE-DMA surface, and pressure is applied tosqueeze out excess TMPTA monomer. The PFPE-DMA mold is then removed fromthe flat PFPE-DMA surface and pressed against a clean, flatsilicon/silicon oxide wafer and photocured using UV radiation (λ=365 nm)for 10 minutes while under a nitrogen purge. Isolated, poly(TMPTA)features are observed after separation of the PFPE mold and thesilicon/silicon oxide wafer, using scanning electron microscopy (SEM).

4.4 Fabrication of 200-nm Titania Structures for Microelectronics

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, 1 g of Pluronic P123 is dissolved in 12 g of absoluteethanol. This solution was added to a solution of 2.7 mL of concentratedhydrochloric acid and 3.88 mL titanium (IV) ethoxide. Flat, uniform,surfaces are generated by treating a silicon/silicon oxide wafer with“piranha” solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide(aq) solution) and drying. Following this, 50 μL of the sol-gel solutionis then placed on the treated silicon wafer and the patterned PFPE moldplaced on top of it. The substrate is then placed in a molding apparatusand a small pressure is applied to push out excess sol-gel precursor.The entire apparatus is then set aside until the sol-gel precursor hassolidified. Oxide structures will be observed after separation of thePFPE mold and the treated silicon wafer using scanning electronmicroscopy (SEM).

4.5 Fabrication of 200-nm Silica Structures for Microelectronics

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, 2 g of Pluronic P123 is dissolved in 30 g of water and 120 gof 2 M HCl is added while stirring at 35° C. To this solution, add 8.50g of TEOS with stirring at 35° C. for 20 h. Flat, uniform, surfaces aregenerated by treating a silicon/silicon oxide wafer with “piranha”solution (1:1 concentrated sulfuric acid:30% hydrogen peroxide (aq)solution) and drying. Following this, 50 μL of the sol-gel solution isthen placed on the treated silicon wafer and the patterned PFPE moldplaced on top of it. The substrate is then placed in a molding apparatusand a small pressure is applied to push out excess sol-gel precursor.The entire apparatus is then set aside until the sol gel precursor hassolidified. Oxide structures will be observed after separation of thePFPE mold and the treated silicon wafer using scanning electronmicroscopy (SEM).

4.6 Fabrication of 200-nm Europium-Doped Titania Structures forMicroelectronics

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, 1 g of Pluronic P123 and 0.51 g of EuCI₃.6H₂O are dissolvedin 12 g of absolute ethanol. This solution was added to a solution of2.7 mL of concentrated hydrochloric acid and 3.88 mL titanium (IV)ethoxide. Flat, uniform, surfaces are generated by treating asilicon/silicon oxide wafer with “piranha” solution (1:1 concentratedsulfuric acid:30% hydrogen peroxide (aq) solution) and drying. Followingthis, 50 μL of the sol-gel solution is then placed on the treatedsilicon wafer and the patterned PFPE mold placed on top of it. Thesubstrate is then placed in a molding apparatus and a small pressure isapplied to push out excess sol-gel precursor. The entire apparatus isthen set aside until the sol-gel precursor has solidified. Oxidestructures will be observed after separation of the PFPE mold and thetreated silicon wafer using scanning electron microscopy (SEM).

4.7 Fabrication of Isolated “scum free” Features for Microelectronics

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 140-nm lines separated by70 nm. A poly(dimethylsiloxane) mold is used to confine the liquidPFPE-DMA to the desired area. The apparatus is then subjected to UVlight (λ=365 nm) for 10 minutes while under a nitrogen purge. The fullycured PFPE-DMA mold is then released from the silicon master.Separately, TMPTA is blended with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. Flat, uniform, non-wetting surfacescapable of adhering to the resist material are generated by treating asilicon wafer cleaned with “piranha” solution (1:1 concentrated sulfuricacid:30% hydrogen peroxide (aq) solution) and treating the wafer with amixture of an adhesion promoter, (trimethoxysilyl propyl methacrylate)and a non-wetting silane agent (1H, 1H, 2H, 2H-perfluorooctyltrimethoxysilane). The mixture can range from 100% of the adhesionpromoter to 100% of the non-wetting silane. Following this, 50 μL ofTMPTA is then placed on the treated silicon wafer and the patterned PFPEmold placed on top of it. The substrate is then placed in a moldingapparatus and a small pressure is applied to ensure a conformal contactand to push out excess TMPTA. The entire apparatus is then subjected toUV light (λ=365 nm) for ten minutes while under a nitrogen purge.Features are observed after separation of the PFPE mold and the treatedsilicon wafer using atomic force microscopy (AFM) and scanning electronmicroscopy (SEM).

Example 5 Molding of Natural and Engineered Templates

5.1. Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated Using Electron-Beam Lithography

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated using electron beam lithographyby spin coating a bilayer resist of 200,000 MW PMMA and 900,000 MW PMMAonto a silicon wafer with 500-nm thermal oxide, and exposing this resistlayer to an electron beam that is translating in a pre-programmedpattern. The resist is developed in 3:1 isopropanol:methyl isobutylketone solution to remove exposed regions of the resist. A correspondingmetal pattern is formed on the silicon oxide surface by evaporating 5 nmCr and 15 nm Au onto the resist covered surface and lifting off theresidual PMMA/Cr/Au film in refluxing acetone. This pattern istransferred to the underlying silicon oxide surface by reactive ionetching with CF₄/O₂ plasma and removal of the Cr/Au film in aqua regia.(FIG. 31). This master can be used to template a patterned mold bypouring PFPE-DMA containing 1-hydroxycyclohexyl phenyl ketone over thepatterned area of the master. A poly(dimethylsiloxane) mold is used toconfine the liquid PFPE-DMA to the desired area. The apparatus is thensubjected to UV light (λ=365 nm) for 10 minutes while under a nitrogenpurge. The fully cured PFPE-DMA mold is then released from the master.This mold can be used for the fabrication of particles using non-wettingimprint lithography as specified in Particle Fabrication Examples 3.3and 3.4.

5.2 Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated using Photolithography

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated using photolithography by spincoating a film of SU-8 photoresist onto a silicon wafer. This resist isbaked on a hotplate at 95° C. and exposed through a pre-patternedphotomask. The wafer is baked again at 95° C. and developed using acommercial developer solution to remove unexposed SU-8 resist. Theresulting patterned surface is fully cured at 175° C. This master can beused to template a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master, and can be imaged by opticalmicroscopy to reveal the patterned PFPE-DMA mold (see FIG. 32).

5.3 Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated from Dispersed Tobacco Mosaic Virus Particles

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing tobacco mosaicvirus (TMV) particles on a silicon wafer (FIG. 33 a). This master can beused to template a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. The morphology of the mold canthen be confirmed using Atomic Force Microscopy (FIG. 33 b).

5.4. Fabrication of a Perfluoropolvether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated from Block-Copolymer Micelles

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersingpolystyrene-polyisoprene block copolymer micelles on a freshly-cleavedmica surface. This master can be used to template a patterned mold bypouring PFPE-DMA containing 1-hydroxycyclohexyl phenyl ketone over thepatterned area of the master. A poly(dimethylsiloxane) mold is used toconfine the liquid PFPE-DMA to the desired area. The apparatus is thensubjected to UV light (λ=365 nm) for 10 minutes while under a nitrogenpurge. The fully cured PFPE-DMA mold is then released from the master.The morphology of the mold can then be confirmed using Atomic ForceMicroscopy (see FIG. 34).

5.5 Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated from Brush Polymers

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing poly(butylacrylate) brush polymers on a freshly-cleaved mica surface. This mastercan be used to template a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. The morphology of the mold canthen be confirmed using Atomic Force Microscopy (FIG. 35).

5.6 Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated from Earthworm Hemoglobin Protein

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing earthwormhemoglobin proteins on a freshly-cleaved mica surface. This master canbe used to template a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. The morphology of the mold canthen be confirmed using Atomic Force Microscopy.

5.7 Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated from Patterned DNA Nanostructures

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing DNAnanostructures on a freshly-cleaved mica surface. This master can beused to template a patterned mold by pouring PFPE-DMA containing1-hydroxycyclohexyl phenyl ketone over the patterned area of the master.A poly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. The morphology of the mold canthen be confirmed using Atomic Force Microscopy.

5.8 Fabrication of a Perfluoropolyether-Dimethacrylate (PFPE-DMA) Moldfrom a Template Generated from Carbon Nanotubes

A template, or “master,” for perfluoropolyether-dimethacrylate(PFPE-DMA) mold fabrication is generated by dispersing or growing carbonnanotubes on a silicon oxide wafer. This master can be used to templatea patterned mold by pouring PFPE-DMA containing 1-hydroxycyclohexylphenyl ketone over the patterned area of the master. Apoly(dimethylsiloxane) mold is used to confine the liquid PFPE-DMA tothe desired area. The apparatus is then subjected to UV light (λ=365 nm)for 10 minutes while under a nitrogen purge. The fully cured PFPE-DMAmold is then released from the master. The morphology of the mold canthen be confirmed using Atomic Force Microscopy.

Example 6 Method of Making Monodisperse Nanostructures having aPlurality of Shapes and Sizes

In some embodiments, the presently disclosed subject matter describes anovel “top down” soft lithographic technique; non-wetting imprintlithography (NoWIL) which allows completely isolated nanostructures tobe generated by taking advantage of the inherent low surface energy andswelling resistance of cured PFPE-based materials.

The presently described subject matter provides a novel “top down” softlithographic technique; non-wetting imprint lithography (NoWIL) whichallows completely isolated nanostructures to be generated by takingadvantage of the inherent low surface energy and swelling resistance ofcured PFPE-based materials. Without being bound to any one particulartheory, a key aspect of NoWIL is that both the elastomeric mold and thesurface underneath the drop of monomer or resin are non-wetting to thisdroplet. If the droplet wets this surface, a thin scum layer willinevitably be present even if high pressures are exerted upon the mold.When both the elastomeric mold and the surface are non-wetting (i.e. aPFPE mold and fluorinated surface) the liquid is confined only to thefeatures of the mold and the scum layer is eliminated as a seal formsbetween the elastomeric mold and the surface under a slight pressure.Thus, the presently disclosed subject matter provides for the first timea simple, general, soft lithographic method to produce nanoparticles ofnearly any material, size, and shape that are limited only by theoriginal master used to generate the mold.

Using NoWIL, nanoparticles composed of 3 different polymers weregenerated from a variety of engineered silicon masters. Representativepatterns include, but are not limited to, 3-μm arrows (see FIG. 11),conical shapes that are 500 nm at the base and converge to <50 nm at thetip (see FIG. 12), and 200-nm trapezoidal structures (see FIG. 13).Definitive proof that all particles were indeed “scum-free” wasdemonstrated by the ability to mechanically harvest these particles bysimply pushing a doctor's blade across the surface. See FIGS. 20 and 22.

Polyethylene glycol (PEG) is a material of interest for drug deliveryapplications because it is readily available, non-toxic, andbiocompatible. The use of PEG nanoparticles generated by inversemicroemulsions to be used as gene delivery vectors has previously beenreported. K. McAllister et al., Journal of the American Chemical Society124, 15198-15207 (Dec. 25, 2002). In the presently disclosed subjectmatter, NoWIL was performed using a commercially availablePEG-diacrylate and blending it with 1 wt % of a photoinitiator,1-hydroxycyclohexyl phenyl ketone. PFPE molds were generated from avariety of patterned silicon substrates using a dimethacrylatefunctionalized PFPE oligomer (PFPE DMA) as described previously. See J.P. Rolland, E. C. Hagberg, G. M. Denison, K. R. Carter, J. M. DeSimone,Angewandte Chemie-International Edition 43, 5796-5799 (2004). In oneembodiment, flat, uniform, non-wetting surfaces were generated by usinga silicon wafer treated with a fluoroalkyl trichlorosilane or by castinga film of PFPE-DMA on a flat surface and photocuring. A small drop ofPEG diacrylate was then placed on the non-wetting surface and thepatterned PFPE mold placed on top of it. The substrate was then placedin a molding apparatus and a small pressure was applied to push out theexcess PEG-diacrylate. The entire apparatus was then subjected to UVlight (A=365 nm) for ten minutes while under a nitrogen purge. Particleswere observed after separation of the PFPE mold and flat, non-wettingsubstrate using optical microscopy, scanning electron microscopy (SEM),and atomic force microscopy (AFM).

Poly(lactic acid) (PLA) and derivatives thereof, such aspoly(lactide-co-glycolide) (PLGA), have had a considerable impact on thedrug delivery and medical device communities because it isbiodegradable. See K. E. Uhrich, S. M. Cannizzaro, R. S. Langer, K. M.Shakesheff, Chemical Reviews 99, 3181-3198 (November 1999); A. C.Albertsson, I. K. Varma, Biomacromolecules 4, 1466-1486(November-December 2003). As with PEG-based systems, progress has beenmade toward the fabrication of PLGA particles through various dispersiontechniques that result in size distributions and are strictly limited tospherical shapes. See C. Cui, S. P. Schwendeman, Langmuir 34, 8426(2001).

The presently disclosed subject matter demonstrates the use of NoWIL togenerate discrete PLA particles with total control over shape and sizedistribution. For example, in one embodiment, one gram of(3S)-cis-3,6-dimethyl-1,4-dioxane-2,5-dione was heated above its meltingtemperature to 110° C. and ˜20 μL of stannous octoate catalyst/initiatorwas added to the liquid monomer. A drop of the PLA monomer solution wasthen placed into a preheated molding apparatus which contained anon-wetting flat substrate and mold. A small pressure was applied aspreviously described to push out excess PLA monomer. The apparatus wasallowed to heat at 110° C. for 15 h until the polymerization wascomplete. The PFPE-DMA mold and the flat, non-wetting substrate werethen separated to reveal the PLA particles.

To further demonstrate the versatility of NoWIL, particles composed of aconducting polymer polypyrrole (PPy) were generated. PPy particles havebeen formed using dispersion methods, see M. R. Simmons, P. A. Chaloner,S. P. Armes, Langmuir 11, 4222 (1995), as well as “lost-wax” techniques,see P. Jiang, J. F. Bertone, V. L. Colvin, Science 291, 453 (2001).

The presently disclosed subject matter demonstrates for the first time,complete control over shape and size distribution of PPy particles.Pyrrole is known to polymerize instantaneously when in contact withoxidants such as perchloric acid. Dravid et al. has shown that thispolymerization can be retarded by the addition of tetrahydrofuran (THF)to the pyrrole. See M. Su, M. Aslam, L. Fu, N. Q. Wu, V. P. Dravid,Applied Physics Letters 84, 4200-4202 (May 24, 2004).

The presently disclosed subject matter takes advantage of this propertyin the formation of PPy particles by NoWIL. For example, 50 μL of a 1:1v/v solution of THF:pyrrole was added to 50 μL of 70% perchloric acid. Adrop of this clear, brown solution (prior to complete polymerization)into the molding apparatus and applied pressure to remove excesssolution. The apparatus was then placed into the vacuum oven overnightto remove the THF and water. PPy particles were fabricated with goodfidelity using the same masters as previously described.

Importantly, the materials properties and polymerization mechanisms ofPLA, PEG, and PPy are completely different. For example, while PLA is ahigh-modulus, semicrystalline polymer formed using a metal-catalyzedring opening polymerization at high temperature, PEG is a malleable,waxy solid that is photocured free radically, and PPy is a conductingpolymer polymerized using harsh oxidants. The fact that NoWIL can beused to fabricate particles from these diverse classes of polymericmaterials that require very different reaction conditions underscoresits generality and importance.

In addition to its ability to precisely control the size and shape ofparticles, NoWIL offers tremendous opportunities for the facileencapsulation of agents into nanoparticles. As described in Example3-14, NoWIL can be used to encapsulate a 24-mer DNA strand fluorescentlytagged with CY-3 inside the previously described 200 nm trapezoidal PEGparticles. This was accomplished by simply adding the DNA to themonomer/water solution and molding them as described. We were able toconfirm the encapsulation by observing the particles using confocalfluorescence microscopy (see FIG. 28). The presently described approachoffers a distinct advantage over other encapsulation methods in that nosurfactants, condensation agents, and the like are required.Furthermore, the fabrication of monodisperse, 200 nm particlescontaining DNA represents a breakthrough step towards artificialviruses. Accordingly, a host of biologically important agents, such asgene fragments, pharmaceuticals, oligonucleotides, and viruses, can beencapsulated by this method.

The method also is amenable to non-biologically oriented agents, such asmetal nanoparticles, crystals, or catalysts. Further, the simplicity ofthis system allows for straightforward adjustment of particleproperties, such as crosslink density, charge, and composition by theaddition of other comonomers, and combinatorial generation of particleformulations that can be tailored for specific applications.

Accordingly, NoWIL is a highly versatile method for the production ofisolated, discrete nanostructures of nearly any size and shape. Theshapes presented herein were engineered non-arbitrary shapes. NoWIL caneasily be used to mold and replicate non-engineered shapes found innature, such as viruses, crystals, proteins, and the like. Furthermore,the technique can generate particles from a wide variety of organic andinorganic materials containing nearly any cargo. The method issimplistically elegant in that it does not involve complex surfactantsor reaction conditions to generate nanoparticles. Finally, the processcan be amplified to an industrial scale by using existing softlithography roller technology, see Y. N. Xia, D. Qin, G. M. Whitesides,Advanced Materials 8, 1015-1017 (December 1996), or silk screen printingmethods.

Example 7 Synthesis of Functional Perfluoropolyethers

7.1. Synthesis of Krytox® (DuPont, Wilmington, Del., United States ofAmerica) Diol to be Used as a Functional PFPE

7.2 Synthesis of Krytox® (DuPont, Wilmington, Del., United States ofAmerica) Diol to be Used as a Functional PFPE

7.3 Synthesis of Krytox® (DuPont, Wilmington, Del., United States ofAmerica) Diol to be Used as a Functional PFPE

7.4 Example of Krytox® (DuPont, Wilmington, Del., United States ofAmerica) Diol to be Used as a Functional PFPE

7.5 Synthesis of a Multi-Arm PFPE Precursor

wherein, X includes, but is not limited to an isocyanate, an acidchloride, an epoxy, and a halogen; R includes, but is not limited to anacrylate, a methacrylate, a styrene, an epoxy, and an amine; and thecircle represents any multifunctional molecule, such a cyclic compound.PFPE can be any perfluoropolyether material as described herein,including, but not limited to a perfluoropolyether material including abackbone structure as follows:

7.6 Synthesis of a Hyperbranched PFPE Precursor

wherein, PFPE can be any perfluoropolyether material as describedherein, including, but not limited to a perfluoropolyether materialincluding a backbone structure as follows:

Example 8 Variability of Congruent Particle Fabrication

8.1 Fabrication of 200 nm Trapezoidal Particles from Various MatrixMaterials

To demonstrate the utility and flexibility of PRINT, shape specificorganic particles composed of three different materials were generatedfrom a commercially available silicon template (FIG. 49A) that iscomposed of a 2 dimensional array of 200 nm trapezoids. Elastomeric PFPEreplica molds of the silicon master templates were generated by pouringa PFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over the silicon substrate patterned with 200-nm trapezoidalshapes. A poly(dimethylsiloxane) perimeter mold is used to confine theliquid PFPE-DMA to a desired area. The apparatus was then subjected toUV light (λ=365 nm) for 10 minutes while under a nitrogen purge. Thefully cured PFPE-DMA mold was then released from the silicon master.This process was repeated to obtain several molds of the same master.

To fabricate monodisperse PLA particles using the PRINT™ process, onegram of (3S)-cis-3,6-dimethyl-1,4-dioxane-2,5-dione (melting point 92°C.) was heated to 110° C. and approximately 20 μL of stannous octoatecatalyst/initiator is added to the liquid monomer. Flat, uniform,non-wetting surfaces are generated by treating a silicon wafer cleanedwith “piranha” solution (1:1 concentrated sulfuric acid: 30% hydrogenperoxide (aq) solution) with trichloro(1H, 1H, 2H, 2H-perfluorooctyl)silane via vapor deposition in a desiccator for 20 minutes. Followingthis, 50 μL of molten Lactic acid containing catalyst is then placed onthe treated silicon wafer preheated to 110° C. and the patterned PFPEmold is placed on top of it. A small pressure is applied to the top ofthe mold with a planar surface to push out excess monomer. The entireapparatus is then placed in an oven at 110° C. for 15 hours. Afterpolymerization was achieved, the PFPE mold and the flat, nonwettingsubstrate were separated to reveal monodisperse 200 nm trapezoidalparticles (FIG. 49B).

To further demonstrate the versatility and breadth of the PRINTtechnique, we chose to generate specifically shaped particles of 200 nmtrapezoids from poly(pyrrole) (PPy). PPy has been used in a variety ofapplications, ranging from electronic devices and sensors to cellscaffolds. We fabricated PPy particles via one-step polymerization usingthe following method: flat, uniform, non-wetting surfaces are generatedby treating a silicon wafer cleaned with “piranha” solution (1:1concentrated sulfuric acid: 30% hydrogen peroxide (aq) solution) withtrichloro(1H, 1H, 2H, 2H-perfluorooctyl) silane via vapor deposition ina desiccator for 20 minutes. Separately, 50 μL of a 1:1 v:v solution oftetrahydrofuran:pyrrole is added to 50 μL of 70% perchloric acid (aq). Aclear, homogenous, brown solution quickly forms and develops into black,solid, polypyrrole in 15 minutes. A drop of this clear, brown solution(prior to complete polymerization) is placed onto a treated siliconwafer, the PFPE mold is placed on top, and pressure is applied with aplanar surface to remove excess solution. The apparatus is then placedinto a vacuum oven for 15 h to remove the THF and water. Particles areobserved using scanning electron microscopy (SEM) (see FIG. 49C) afterrelease of the vacuum and separation of the PFPE mold and the treatedsilicon wafer.

Trapezoidal trimethylopropane triacrylate (TMPTA) particles were alsogenerated using a photopolymerization technique. TMPTA is blended with 1wt % of a photoinitiator, 1-hydroxycyclohexyl phenyl ketone. Uniform,non-wetting surfaces are generated by pouring a PFPE-dimethacrylate(PFPE-DMA) containing 1-hydroxycyclohexyl phenyl ketone over a siliconwafer. The wafer was then subjected to UV light (λ=365 nm) for 10minutes while under a nitrogen purge. The fully cured PFPE-DMA substratewas then released from the silicon master. Following this, 50 μL ofTMPTA is then placed on the PFPE substrate and the patterned PFPE moldplaced on top of it. The substrate is then placed on a flat surface anda small pressure is applied to push out excess TMPTA. The entireapparatus is then subjected to UV light (λ=365 nm) for ten minutes whileunder a nitrogen purge. Particles are observed after separation of thePFPE mold and the treated silicon wafer using scanning electronmicroscopy (SEM). A flat blade was pushed along the surface to gatherthe fabricated 200 nm particles (see FIG. 49D).

Particles of the same unique dimensions made using these three differentpolymerization methods were evaluated using scanning electron microscopyand atomic force microscopy. The NIH Image program was used to measurethe particle dimensions on the micrographs and compare them to images ofthe master template.

8.2 Fabrication of PEG Particles of Different Shapes and Sizes

Poly(ethylene glycol) (PEG) is a material of tremendous interest to thebiotechnology community due to its commercial availability, nontoxicnature, and biocompatibility. Here, the PRINT was utilized to producemonodisperse, micro- and nanometer scale PEG particles in a variety ofshapes by molding a PEG-diacrylate liquid monomer followed by roomtemperature photopolymerization. Because the morphology of the particlesis controlled by the master, it is possible to generate complexparticles on a variety of length scales.

A patterned perfluoropolyether (PFPE) molds are generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with the desired shape. Thesilicon masters used include: 200 nm trapezoidal features (FIG. 50A);200 nm×800 nm bars (FIG. 50B); 500 nm conical features that are <50 nmat the tip (FIG. 50C); 3 μm arrows (FIG. 50D); 10 μm boomerangs (FIG.50E); and 600 nm cylinders (FIG. 50F). The master coated with uncuredPFPE was then subjected to UV light (λ=365 nm) for 10 minutes whileunder a nitrogen purge. The fully cured PFPE-DMA mold was then easilyreleased from the silicon master by peeling. Separately, a poly(ethyleneglycol) (PEG) diacrylate (n=9) is blended with 1 wt % of aphotoinitiator, 1-hydroxycyclohexyl phenyl ketone. Uniform, non-wettingsurfaces are generated by pouring a PFPE-dimethacrylate (PFPE-DMA)containing 1-hydroxycyclohexyl phenyl ketone over a silicon wafer. Thewafer was then subjected to UV light (λ=365 nm) for 10 minutes whileunder a nitrogen purge. The fully cured PFPE-DMA substrate was thenreleased from the silicon master. Following this, 50 μL of PEGdiacrylate is then placed on the PFPE film and the patterned PFPE moldplaced on top of it. The substrate is then placed on a flat surface anda small pressure is applied to push out excess PEG-diacrylate. Thepressure used was at least about 100 N/cm². The entire apparatus wasthen subjected to UV light (λ=365 nm) for ten minutes while under anitrogen purge. Arrays of particles of different shapes and sizes areobserved after separation of the PFPE mold and the treated silicon waferusing scanning electron microscopy (SEM). (See FIGS. 50A-50F).

Confirmation of the structural similarity between the silicon master andreplicate PEG particles was confirmed via atomic force microscopy (AFM)and scanning electron microscopy (SEM). Atomic Force Microscopy wasperformed on a Nanoscope IIIa/Multimode AFM in tapping mode. Dynamiclight scattering (DLS) is performed on particles suspended in phosphatebuffered saline solution (PBS) to look for aggregation. This techniqueis designed for spherical particles; however, we can use the valuesempirically to look for large aggregates (some non-uniformity in sizewill be seen at a scale smaller than that of the particle diameter dueto the non-spherical shapes of the particles) An example DLS trace isgiven in FIG. 51, with the value measured for the particle size as0.62±0.2 μm. The line indicates monodispersity of the particles, with noaggregation occurring.

8.3 Utilizing PRINT Technology to Create Free-Flowing Particles,Particles on a Scum Layer, and Particles on a Film

The PRINT technology can be used to generate a variety of productshaving varying forms, including free flowing particles and particles inan array on a film. The following example shows our ability to makepoly(ethylene glycol) (PEG) based particles free flowing, as an array ona PEG film, and as an array on a different polymer film.

Free-flowing Particles: A patterned perfluoropolyether (PFPE) mold wasgenerated by pouring a PFPE-dimethacrylate (PFPE-DMA) containing1-hydroxycyclohexyl phenyl ketone over a silicon substrate patternedwith 200 nm tall×200 nm diameter cylinders. The PFPE-DMA covered masterwas then subjected to UV light (λ=365 nm) for 3 minutes while under anitrogen purge. The fully cured PFPE-DMA mold was then released from thesilicon master. Separately, a mixture of 790 mg trimethylolpropaneethoxylate triacrylate, 200 mg polyethylene glycol carbonylimidizolemonomethacrylate, and 10 mg α-α-diethoxyacetophenone was prepared. Thismixture was spotted directly onto the patterned PFPE-DMA mold andcovered with an unpatterned polyethylene (PE) film. The monomer mixturewas pressed between the two polymer sheets, and then the PE sheet wasslowly peeled from the patterned PFPE-DMA to remove any excess monomersolution from the surface of the PFPE-DMA mold. The mold was thensubjected to UV light (λ=365 nm) for 2 minutes while maintaining anitrogen purge. The particles were harvested by placing 2 mL of DMSO onthe mold and scrapping the surface with a glass slide. The particlesuspension was transferred to a scintillation vial. One drop of thesuspension was placed on a SEM stub and the solvent was allowed toevaporate. The stub was coated with approximately 10 angstroms of goldand imaged with SEM (FIG. 52A).

Particles on a PEG film: A patterned perfluoropolyether (PFPE) mold isgenerated by pouring a PFPE-dimethacrylate (PFPE-DMA) containing1-hydroxycyclohexyl phenyl ketone over a 6 inch silicon substratepatterned with 200-nm cylindrical shapes. The substrate is thensubjected to UV light (A =365 nm) for 10 minutes while under a nitrogenpurge. The fully cured PFPE-DMA mold is then released from the siliconmaster. Separately, a solution of 30:70 PEG monomethacrylate:PEGdiacrylate is formulated with 1 wt % photoinitiator. Following this, 200μL of this PEG solution is then placed on an untreated silicon wafer andthe patterned PFPE mold placed on top of it. The substrate is thenplaced on a flat substrate and a small pressure is applied to push outexcess PEG solution. The entire apparatus is then subjected to UV light(λ=365 nm) for ten minutes while under a nitrogen purge. PEG particlesconnected by a PEG film will be observed after separation of the PFPEmold and the silicon wafer using scanning electron microscopy. Dragginga blade across the surface yields a rolled up film as shown in FIG. 52B.

Particles on a cyanoacrylate film: A patterned perfluoropolyether (PFPE)mold is generated by pouring a PFPE-dimethacrylate (PFPE-DMA) containing2,2-diethoxyacetophenone over a silicon substrate patterned with 200 nmcylindrical shapes. The apparatus is then subjected to a nitrogen purgefor 10 minutes before the application of UV light (λ=365 nm) for 10minutes while under a nitrogen purge. The fully cured PFPE-DMA mold isthen released from the silicon master. Separately, a poly(ethyleneglycol) (PEG) diacrylate (n=9) is blended with 28 wt % PEG methacrylate(n=9), 2 wt % azobisisobutyronitrile (AIBN), and 0.25 wt % rhodaminemethacrylate. Flat, uniform, non-wetting surfaces are generated bycoating a glass slide with PFPE-dimethacrylate (PFPE-DMA) containing2,2-diethoxyacetophenone. The slide is then subjected to a nitrogenpurge for 10 minutes, then UV light is applied (λ=365 nm) while under anitrogen purge. The flat, fully cured PFPE-DMA substrate is releasedfrom the slide. Following this, 0.1 mL of the monomer blend is evenlyspotted onto the flat PFPE-DMA surface and then the patterned PFPE-DMAmold placed on top of it. The surface and mold are then placed in amolding apparatus and a small amount of pressure is applied to removeany excess monomer solution. The entire apparatus is purged withnitrogen for 10 minutes, then subjected to UV light (λ=365 nm) for 10minutes while under a nitrogen purge. Neutral PEG nanoparticles areobserved after separation of the PFPE-DMA mold and substrate usingscanning electron microscopy (SEM). A thin layer of cyanoacrylatemonomer is sprayed onto the PFPE-DMA mold filled with particles. ThePFPE-DMA mold is immediately placed onto a glass slide and thecyanoacrylate is allowed to polymerize in an anionic fashion for oneminute. The mold is removed and the particles are embedded in theadhesive layer (see FIG. 52C).

8.4 Identification of PRINT Particles Using Nano-Scale “Defects”

The PRINT process inherently introduces structural features from thesilicon masters that are transferred to the mold and subsequently to theparticles during PRINT fabrication. Here, a Bosch-type etch is used toprocess a master which introduces a recognizable pattern (“Bosch etchlines”) on the sidewalls of individual particles. Bosch etching is oneof many techniques used to fabricate wafers, most of which leaveresidual “defects” on the sidewalls of the features or surface. FIGS.53A and 53B shows distinct particles derived from the masters that showa similar sidewall pattern resulting from the specific Bosch-type etchprocess used on the master. In this case, this pattern can be recognizedusing SEM imaging and identifies these particles as originating from thesame master.

A patterned perfluoropolyether (PFPE) mold is generated by pouring aPFPE-dimethacrylate (PFPE-DMA) containing 1-hydroxycyclohexyl phenylketone over a silicon substrate patterned with 3-μm cubical shapes at a1 μm depth. The substrate is then subjected to a nitrogen purge for 10minutes, then UV light (λ=365 nm) is applied for 10 minutes while undera nitrogen purge. The fully cured PFPE-DMA mold is then released fromthe silicon master. A PFPE-DMA mold is made from a master patterned with2 μm deep cubical shapes Separately, TMPTA is blended with 1 wt % of aphotoinitiator, 1-hydroxycyclohexyl phenyl ketone. Flat, uniform,non-wetting surfaces are generated by coating a glass slide withPFPE-DMA containing 1-hydroxycyclohexyl phenyl ketone. The slide is thensubjected to a nitrogen purge for 10 minutes, then UV light (λ=365 nm)is applied for 10 minutes while under a nitrogen purge. The flat, fullycured PFPE-DMA substrate is released from the slide. Following this, 0.1mL of TMPTA is then placed on the flat PFPE-DMA substrate and thepatterned PFPE mold placed on top of it. The substrate is then placed ina molding apparatus and a small pressure is applied to push out excessTMPTA. The entire apparatus is then purged with nitrogen for 10 minutes,then subjected to UV light (λ=365 nm) for 10 minutes while under anitrogen purge. TMPTA particles are observed after separation of thePFPE-DMA mold and substrate using optical microscopy. A drop ofn-vinyl-2-pyrrolidone containing 5% photoinitiator, 1-hydroxycyclohexylphenyl ketone, is placed on a clean glass slide. The PFPE-DMA moldcontaining particles is placed patterned side down on then-vinyl-2-pyrrolidone drop. The slide is subjected to a nitrogen purgefor 5 minutes, then UV light (λ=365 nm) is applied for 5 minutes whileunder a nitrogen purge. The slide is removed, and the mold is peeledaway from the polyvinyl pyrrolidone and particles. Particles on thepolyvinyl pyrrolidone were observed with optical microscopy. Thepolyvinyl pyrrolidone film containing particles was dissolved in water.Dialysis was used to remove the polyvinyl pyrrolidone, leaving anaqueous solution containing TMPTA particles. Samples dispersions fromthe 1 μm and 2 μm deep master are dropped on an SEM stub and the waterallowed to evaporate in a vacuum oven. The particles were coated with˜10 Å gold-palladium and imaged with SEM (FIGS. 53A and 53B).

What is claimed is:
 1. A delivery device, comprising: a dissolvablesubstrate of a first material, wherein the dissolvable substrate isdissolvable in biological tissues; and a plurality of substantiallymonodisperse particles configured from a second material; each particleof the plurality comprising: a predetermined, non-sphericalthree-dimensional geometric solid shape; parallel lateral surfaces andparallel top and bottom surfaces in cross-section; and a maximumcross-sectional dimension of less than about 10 micrometers; wherein theplurality of substantially monodisperse particles are coupled with thedissolvable substrate in a substantially predetermined orientation. 2.The delivery device of claim 1, wherein the maximum cross-sectionaldimension is between about 5 nanometers and about 5 micrometers.
 3. Thedelivery device of claim 1, wherein the maximum cross-sectionaldimension is between about 10 nanometers and about 2 micrometers.
 4. Thedelivery device of claim 1, wherein the maximum cross-sectionaldimension is between about 10 nanometers and about 1 micrometer.
 5. Thedelivery device of claim 1, wherein the maximum cross-sectionaldimension is less than about 1 micrometer.
 6. The delivery device ofclaim 1, wherein the maximum cross-sectional dimension is less thanabout 750 nanometers.
 7. The delivery device of claim 1, wherein themaximum cross-sectional dimension is less than about 500 nanometers. 8.The delivery device of claim 1, wherein the maximum cross-sectionaldimension is less than about 300 nanometers.
 9. The delivery device ofclaim 1, wherein the maximum cross-sectional dimension is less thanabout 250 nanometers.
 10. The delivery device of claim 1, wherein themaximum cross-sectional dimension is less than about 200 nanometers. 11.The delivery device of claim 1, wherein the maximum cross-sectionaldimension is less than about 150 nanometers.
 12. The delivery device ofclaim 1, wherein the maximum cross-sectional dimension is less thanabout 100 nanometers.
 13. The delivery device of claim 1, wherein thesecond material comprises an organic material.
 14. The delivery deviceof claim 1, wherein the second material comprises an imaging agent. 15.The delivery device of claim 1, wherein the second material comprises adrug.
 16. The delivery device of claim 1, wherein the second materialcomprises a treatment agent.
 17. The delivery device of claim 1, whereinthe second material comprises an antibiotic.
 18. The delivery device ofclaim 1, wherein the second material comprises biologic material. 19.The delivery device of claim 1, wherein the second material comprises asoluble material.
 20. The delivery device of claim 1, wherein the secondmaterial comprises a biodegradable material.
 21. The delivery device ofclaim 1, wherein the second material comprises a hydrophilic material.22. The delivery device of claim 1, wherein the second materialcomprises a hydrophobic material.
 23. The delivery device of claim 1,wherein the second material comprises an inorganic material.
 24. Thedelivery device of claim 1, wherein the second material comprises apolymer material.
 25. The delivery device of claim 1, wherein the secondmaterial comprises a small molecule.
 26. The delivery device of claim 1,wherein the first material and the second material comprise the samecomposition.
 27. The delivery device of claim 1, wherein said deliverydevice has a modulus from about 0.1 MPa to about 500 MPa.
 28. Thedelivery device of claim 1, wherein delivery device has a modulus ofabout 1 MPa to about 100 MPa.
 29. The delivery device of claim 1,wherein the second material comprises a porogen.
 30. The delivery deviceof claim 1, wherein the second material comprises a thermoplasticmaterial.
 31. The delivery device of claim 1, wherein the dissolvablesubstrate comprises a thickness of about twice a dimension of a particleof the plurality of particles.
 32. The delivery device of claim 1,wherein the dissolvable substrate comprises a thickness of about equal adimension of a particle of the plurality of particles.
 33. The deliverydevice of claim 1, wherein the dissolvable substrate comprises athickness of about one half a dimension of a particle of the pluralityof particles.
 34. The delivery device of claim 1, further comprisingparticles of more than one size coupled with the dissolvable substrate.35. The delivery device of claim 1, further comprising particles of morethan one shape coupled with the dissolvable substrate.
 36. The deliverydevice of claim 1, further comprising an agent configured to couple theplurality of substantially monodisperse particles to the dissolvablesubstrate, wherein the agent is selected from the group consisting ofcovalent bonding, ionic bonding, electrostatic binding, surface energy,hydrogen bonding, van der Waals forces, other intra- and inter-molecularforces, adhesives, and a magnetic force.
 37. A method of fabricating adelivery device of claim 1, comprising: placing a first material into aplurality of recesses in a polymer mold, wherein each recess is lessthan about 10 micrometers in a maximum cross-sectional dimension;hardening the first material to form a plurality of substantiallymonodisperse particles, each particle of the plurality comprising apredetermined, non-spherical geometric solid shape substantiallycorresponding to the recess and parallel lateral surfaces and paralleltop and bottom surfaces in cross-section; removing the plurality ofsubstantially monodisperse particles from the recesses; and coupling theplurality of substantially monodisperse particles with a dissolvablesubstrate, wherein the dissolvable substrate is dissolvable inbiological tissues; and the plurality of substantially monodisperseparticles are arranged in a substantially predetermined orientation. 38.The method of claim 37, wherein the maximum cross-sectional dimension isbetween about 5 nanometers and about 5 micrometers.
 39. The method ofclaim 37, wherein the maximum cross-sectional dimension is between about10 nanometers and about 2 micrometers.
 40. The method of claim 37,wherein the maximum cross-sectional dimension is between about 10nanometers and about 1 micrometer.
 41. The method of claim 37, whereinthe maximum cross-sectional dimension is less than about 1 micrometer.42. The method of claim 37, wherein the maximum cross-sectionaldimension is less than about 750 nanometers.
 43. The method of claim 37,wherein the maximum cross-sectional dimension is less than about 500nanometers.
 44. The method of claim 37, wherein the maximumcross-sectional dimension is less than about 300 nanometers.
 45. Themethod of claim 37, wherein the maximum cross-sectional dimension isless than about 250 nanometers.
 46. The method of claim 37, wherein themaximum cross-sectional dimension is less than about 200 nanometers. 47.The method of claim 37, wherein the maximum cross-sectional dimension isless than about 150 nanometers.
 48. The method of claim 37, wherein themaximum cross-sectional dimension is less than about 100 nanometers. 49.The delivery device of claim 1, wherein said plurality of substantiallymonodisperse particles are arranged in a substantially ordered array.50. The delivery device of claim 1, wherein said dissolvable substrateis a sugar-based dissolvable film.
 51. The delivery device of claim 50,wherein said film is a sugar sheet comprising pullulan,hydroxypropylmethyl cellulose, hydroxyethyl cellulose, hydroxypropylcellulose, polyvinyl pyrrolidone, carboxymethyl cellulose, polyvinylalcohol, sodium alginate, polyethylene glycol, xanthan gum, tragacanthgum, guar gum, acacia gum, arabic gum, polyacrylic acid,methylmethacrylate copolymer, carboxyvinyl polymer, amylose, highamylose starch, hydroxypropylated high amylose starch, dextrin, pectin,chitin, chitosan, levan, elsinan, collagen, gelatin, zein, gluten, soyprotein isolate, whey protein isolate and/or casein, or combinationsthereof.